{"title":"IC50 \u0026 Dose-Response","description":null,"products":[{"product_id":"tr-fret-cereblon-4c-binding-assay-kit-bht20700004","title":"TR-FRET Cereblon-4C Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eE3 ubiquitin ligases are a large family of enzymes that catalyze the critical final step in the ubiquitination \ncascade. Cereblon (CRBN) serves as the substrate-recognition component of the E3 ubiquitin ligase \n\ncomplex CRBN\/DDB1\/CUL4A\/RBX1 (CRBN-4C). CRBN is one of the most widely utilized E3 ligases \nin the design of PROTACs (Proteolysis-Targeting Chimeras) for targeted protein degradation (TPD) in \n\ndrug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe TR-FRET CRBN Binding Assay Kit is designed to measure the binding affinity between CRBN and \nits ligands. The kit includes Tag1-CRBN-4C (a complex of CRBN\/CUL4A\/DDB1\/RBX1), a terbium-\nlabeled anti-Tag1 antibody, and a fluorescently labeled CRBN ligand, thalidomide. When CRBN binds \nto thalidomide, the terbium donor (on the anti-Tag1 antibody) is brought into close proximity to the \nfluorescent acceptor (fluorophore-labeled thalidomide), resulting in fluorescence resonance energy \ntransfer (FRET). The binding interaction can be quantitatively measured as an HTRF signal, calculated \nfrom the ratio of the emission intensities at 665 nm (acceptor) and 620 nm (donor). If a test compound \ncompetes with thalidomide for CRBN binding, the HTRF signal will decrease, indicating inhibition of \nCRBN–thalidomide interaction.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that bind to Cereblon for drug discovery. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e272625-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-4C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003eFluorescence-labeled Thalidomide\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare CRBN-4C solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare FL-Thalidomide solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate sample HTRF signal of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare CRBN-4C solution Thaw CRBN-4C protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: CRBN-4C protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the CRBN-4C protein 20-fold (1µL CRBN-4C + 19 µL assay buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of assay buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of assay buffer to each of negative and positive control wells. If the compound is diluted in 10% DMSO, add 2 µl of assay buffer containing 10% DMSO to each of\tnegative and positive control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare FL-Thalidomide solution Dilute FL-Thalidomide 125-fold (1 µL FL-Thalidomide + 124 µL of assay buffer). Add 4 µl of diluted FL-Thalidomide solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag1 antibody 1:200 in assay buffer. For example: 1 µl of Terbium- labeled anti-Tag1 antibody + 199 µl of assay buffer. Add 10 µl of this dye mixture to each well. -4510 or 858453-5700 Fax: 855-898-3979 3\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Incubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eUbiquitin-Proteasome \/ Targeted Protein Degradation\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology; Hematology; PROTAC Drug Discovery\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238301983085,"sku":"272625","price":1299.0,"currency_code":"USD","in_stock":false}]},{"product_id":"sars-cov-2-nucleocapsid-protein-binding-kit-for-rabbit-antibody-bht20700029","title":"SARS-CoV-2 Nucleocapsid Protein Binding Kit (For rabbit antibody)","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eSARS-CoV-2 Nucleocapsid protein (NP) is one of the core components of SARS-CoV-2 virus. It forms \na complex with viral genomic RNA in a helical symmetrical structure and plays a key role in the process \nof virus replication and assembly. Since NP is abundantly expressed during infection, it can be used \nas an important diagnostic marker for COVID-19 and also can be used as a potential drug target or \ndeveloping vaccines.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe SARS-CoV-2 Nucleocapsid protein (NP) Binding kit is a TR-FRET based assay, that is designed \nto detect binding status of NP to the test antibody. Terbium-labeled anti-Tag1 antibody serves as \nfluorescence donor, that binds to the His-Tagged NP. If a test rabbit antibody binds to NP, fluorescence-\nlabeled anti-rabbit antibody (fluorescence acceptor) will be brought in close proximity with the \nfluorescence donor. Excitation of Terbium (340 nm) generates fluorescence resonance energy transfer \n(FRET) to the fluorescence-labeled acceptor, which consequently fluoresces at 665 nm (figure below). \nThus, the test antibody binding to NP can be quantitively measured by calculation of the fluorescent \nratio of 665 nm\/620 nm.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of antibodies that bind to NP.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eAurora Biolabs LLC, San Diego, CA 92121, USA; www.aurorabiolabs.com; SARS-CoV-2 Nucleocapsid Protein Binding Kit (for rabbit antibody) A HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e2x Assay Buffer\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003eCustomer Test anti-NP-rabbit antibody (to be tested antibody)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare SARS-CoV-2 Nucleocapsid protein Dilute SARS-CoV-2 Nucleocapsid protein (NP) 1,500-fold with 1X DTT-containing assay buffer. For example: 1 µl of NP + 1,499 µl of 1X DTT-containing assay buffer. Add 5 µl of diluted NP protein to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare Antibody solution\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag1 Ab and fluorescence-labeled anti-rabbit antibody 1:200 in 1X DTT- containing assay buffer. For example: 1 µl of Terbium-labeled anti-Tag1 Ab + 1 µl of fluorescence- labeled anti-rabbit antibody + 198 µl of 1X DTT-containing assay buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Incubate the reaction at room temperature for 1 hour.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample signal. Calculate percentage activity \n\nAurora Biolabs LLC, San Diego, CA 92121, USA; www. SARS-CoV-2 Nucleocapsid Protein Binding Kit \n(for rabbit antibody) In the absence of the compound (positive control), the sample signal (P) is defined as 100% activity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% activity. The \npercent activity in the presence of each compound is calculated according to the following \nequation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence of the \ncompound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e Nucleocapsid Protein Binding\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302015853,"sku":"728273","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"eif4e-eif4g-binding-assay-kit-bht20700005","title":"eIF4E\/eIF4G Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eEukaryotic translation initiation factor eIF4E, the mRNA cap-binding protein, is considered the rate-\nlimiting factor in translation. It plays an important role in cap-dependent translation initiation and \nrecruitment of mRNA to ribosomes. Overexpression of eIF4E has been documented in numerous \nhuman cancers and contributes to transformation, tumorigenesis, and progression of cancers. \nTherefore, eIF4E is an attractive drug target for cancer treatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe eIF4E binding assay kit, a TR-FRET based assay, is designed to screen compounds that bind to \neIF4E. A fluorescence-labelled tracer and the N-terminal tagged full-length human eIF4E\/eIF4G \ncomplex are used in this assay kit. A Terbium-labeled antibody binding to the tag on eIF4E serves as \na fluorescence donor (HTRF donor). The binding of the fluorescence-labeled tracer to the eIF4E brings \nTerbium on the anti-Tag antibody close to the fluorophore on the tracer (HTRF acceptor). Activation of \nthe Terbium results in fluorescence resonance energy transfer (FRET). Thus, the binding status can \nbe quantitively measured by calculating the ratio of the emission fluorescence intensity of the acceptor \n(665 nm) and donor (620 nm). The competitive binding of a non-fluorescence compound will reduce \nthe FRET signal.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that bind to eIF4E\/eIF4G.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eAurora Biolabs LLC, San Diego, CA 92121, USA; www.aurorabiolabs.com; 1 A HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e34343-BK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e16 µL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare assay buffer containing 1 mM DTT\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare eIF4E\/eIF4G solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare fluorescence-labeled tracer\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate the ratio of the fluorescent intensity of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare eIF4E\/eIF4G solution Thaw eIF4E\/eIF4G protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: eIF4E\/eIF4G protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the eIF4E\/eIF4G protein 200-fold (1 µL eIF4E\/eIF4G + 199 µL 1X assay buffer containing DTT). Add 8 µl of diluted protein solution to each positive control wells and inhibitor test wells. Add 8 µl of 1X DTT containing buffer to each of negative control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each negative and positive control wells. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare fluorescence-labeled tracer Thaw the tracer at room temperature. Dilute the tracer 125-fold (1 µL of 1 M tracer + 124 µL 1X assay buffer containing DTT). Add 5 µl of diluted tracer to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag antibody 1:200 (1 µl of Terbium-labeled anti-Tag antibody + 199 µl of 1X DTT-containing assay buffer). Add 5 µl of this dye mixture to each well. Dilute just enough of the antibody for each reaction set. Store remaining undiluted antibody at -80°C. Do not re-use the diluted antibody.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eCap-Dependent Translation Initiation\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (eIF4E-overexpressing tumors)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302048621,"sku":"34343-BK","price":2599.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-wt-nucleotide-exchange-assay-kit-bht20700009","title":"Kras WT Nucleotide Exchange Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway plays an important role in cell proliferation and \n\ndifferentiation. Conversion of Kras from the inactive GDP-bound state to the active GTP-bound state \n\ntriggers the downstream effector and promotes cell growth. RAS genes are frequently mutated in \n\nvarious human tumors. These mutations block the GTPase activity of RAS and lock RAS in the GTP-\n\nbound state, resulting in constitutively active signals through the downstream cascades leading to \n\ncancer cell proliferation.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (wild type, WT) nucleotide exchange assay is a TR-FRET based assay. The assay kit is \n\ndesigned to detect the GTP binding status of wild type Kras in the presence of SOS1, the most-studied \n\nguanine nucleotide exchange factor (GEF) of Kras. The Tag2 Kras in this assay kit is recognized by a \n\nTerbium-labeled anti-Tag2 antibody (HTRF donor). If Kras binds to a fluorescence-labeled GTP (HTRF \n\nacceptor), the donor and the acceptor will be brought in close proximity. Excitation of Terbium (340 nm) \n\ngenerates fluorescence resonance energy transfer (FRET) to the fluorescence-labeled GTP acceptor, \n\nwhich consequently fluoresces at 665 nm (figure below). Thus, GTP binding to Kras can be quantitively \n\nmeasured by calculation of the fluorescent ratio of 665 nm\/620 nm. \n\nAurora Biolabs LLC, San Diego, CA 92121; www.aurorabiolabs.com; \n\nKras (WT) Nucleotide Exchange Assay Kit\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit Kras activation for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-NK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 µL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003e1. Prepare 1X assay buffer containing 1 mM DTT (1X DTT-containing assay buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare SOS1 solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare Kras solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare 1X assay buffer containing 1 mM DTT (1X DTT-containing assay buffer) For example, mix 996 µl distilled water with 1000 µl of 2X assay Buffer (Catalogue number: 5727- NK-B) and 4 µl of 0.5 M DTT. Make only enough 1X DTT-containing assay buffer as needed for the assay. Store the remaining 2X assay buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare SOS1 solution Thaw SOS1 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: SOS1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the SOS1 protein 400-fold (1 µL SOS1 + 399 µL 1X DTT-containing assay buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of 1X DTT-containing assay buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare Kras solution\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled GTP 1:40 in 1X DTT-containing assay buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled GTP + 194 µl of 1X DTT-containing assay buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e17 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302081389,"sku":"5727-4121NK","price":1699.0,"currency_code":"USD","in_stock":false}]},{"product_id":"dna-polymerase-theta-activity-assay-kit-bht20700006","title":"DNA Polymerase Theta Activity Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eDNA polymerase theta (Pol θ) is involved in an end-joining pathway of DNA double-strand breaks. Over \nexpression of Pol θ is found in many cancers, including stomach, colon, breast and lung cancers, and \ncorrelated with poorer patient survival. Because suppression of gene expression of Pol θ results in \nsensitivity of cells to ionizing radiation and some DSB-inducing drugs, Pol θ is a validated anti-cancer drug \ntarget. \n\nDescription \n\nThe Aurora DNA Polymerase Theta activity assay kit is a homogeneous fluorescence-based assay for \nscreening inhibitors that block DNA polymerase activity of DNA Pol θ. \n\n The assay is fast, convenient, and requires just two steps. In the first step, the DNA Pol θ enzyme \nsynthesizes double-stranded DNA using a DNA template in the presence of dNTP. In the second step, a dye \nthat binds to double-stranded DNA is added to the solution resulting in the increase of fluorescence, \nintensity of which can be measured with a fluorescent plate reader at the excitation wavelengths of 495 \nnm and emission wavelengths of 525 nm. \n\nMaterials supplied \n\nCatalogue Number \n362201 \n362204 \n4687 \n362003 \n4930 \n362202 \n\nItem \n2X Assay Buffer \n20 µM DNA template \n10 mM dNTP \nRecombinant DNA Pol θ CTD \nDye solution \nStop solution \nBlack low binding 96 well plate \n\nAmount \n25 mL \n7 µL \n5 µL \n5 µL \n15 µL \n3 mL \n1 \n\nStorage \n-20°C \n-20°C \n-20°C \n-80°C \n-20°C \n-20°C \nRT \n\nMaterials Needed but not supplied \n\nA microplate reader capable of measuring fluorescence at excitation wavelengths of 495 nm and \nemission wavelengths of 525 nm. \n\n1. 0.5 M DTT \n2. Adjustable micro-pipettor \n3. Sterile Tips \n\nStability \n\n12 months if stored under the indicated conditions. \n\nAurora Biolabs LLC, San Diego, CA 92121, USA. www.aurorabiolabs.com, \n\n \n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare 1X buffer containing 1 mM DTT.\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution.\u003c\/li\u003e\n\u003cli\u003ePrepare DNA Pol Theta solution.\u003c\/li\u003e\n\u003cli\u003eAdd the inhibitor solution\u003c\/li\u003e\n\u003cli\u003eIncubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli\u003ePrepare substrate solution\u003c\/li\u003e\n\u003cli\u003eIncubate the plate at 30°C for 1 hour.\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure the fluorescent intensity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution. If the inhibitor compound is dissolved in water, make a solution of the compound 5-fold higher than the final concentration in 1X assay buffer (since you will add 5 µl to the 25 µl reaction). Then make a series of dilutions in 1X assay buffer from this solution to your preferred concentrations. If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 20-fold dilution in 1X assay buffer (at this step, the compound concentration is 5-fold higher than the final concentration and the DMSO concentration is 5%). Then make a series of dilutions in 5% of DMSO from this solution to your preferred concentrations. Since 5 µl of the compound solution will be added to the 25 µl reaction, the final concentration of DMSO in all of reactions is 1%.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare DNA Pol Theta solution. Thaw DNA Pol θ CTD enzyme (catalogue number 362003) on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: DNA Pol θ CTD enzyme is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted enzyme. Dilute DNA Pol Theta enzyme 650-fold (1:650) in 1X assay buffer with 1 mM DTT. Add 10 µl of diluted enzyme solution to each of positive control well and inhibitor test well. Add 1X buffer to each of background well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add the inhibitor solution Add 5 µl of 1X assay buffer to each background well and positive control well if the inhibitor is diluted in 1X buffer. Add 5 µl of 1X assay buffer with 5% DMSO to each of background well and positive control well if the inhibitor is diluted in 1X buffer with 10% DMSO. Add 5 µl of diluted inhibitor solution from Step 2 to each of the inhibitor test well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare substrate solution During the incubation of the enzyme and the inhibitor solution, prepare substrate solution containing 0.125 µM DNA template (dilute from 20 µM DNA template, catalogue number 362004) and 25 µM dNTP (dilute from 10 mM dNTP) in 1X assay buffer. Make only enough solution as need for the assay. Store the remaining 20 µM DNA template and 10 mM dNTP solution to -20°C. Add 10 µl of the substrate solution to each of well, including background wells, positive control wells and the inhibitor test wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the plate at 30°C for 1 hour.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Prepare dye solution Dilute the Dye solution 200-fold in Stop solution (catalogue number 362202). Make only enough solution as need for the assay. Store the remaining Dye solution to -20°C. Add 25 µl the Dye solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Incubate at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Measure the fluorescent intensity Measure the fluorescent intensity at the excitation wavelengths of 495 nm and the emission wavelengths of 525 nm. Positive Control Inhibitor Test\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate percentage activity of the enzyme \n\n% Activity= \n\n(Fp – Fb) – (Fi-Fb) \nFp - Fb \n\nX 100 \n\nWhere Fp refers to fluorescent intensity of the positive control, Fb refers to fluorescent intensity of \nbackground, and Fi refers to fluorescent intensity of the inhibitor. Graph the percentage activity as a function of the inhibitor concentration to determine the IC50 of the test \ninhibitor. No CPD refers to no compound control \n(compound vehicle only).\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e19 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eDNA Double-Strand Break Repair (End-Joining TMEJ)\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (POLQ-overexpressing cancers: breast; lung; colon; stomach)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"96 reactions","offer_id":53238302114157,"sku":"362101-96","price":839.0,"currency_code":"USD","in_stock":false},{"title":"384 reactions","offer_id":53238305554797,"sku":"362101-384","price":1199.0,"currency_code":"USD","in_stock":false}]},{"product_id":"tr-fret-parp1-trapping-assay-kit-bht20700024","title":"TR-FRET PARP1 Trapping Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eTR-FRET PARP1 Trapping Assay Kit \n PARP1 (Poly (ADP-ribose) polymerase 1) is an abundant member of the PARP family and plays a \ncrucial role in DNA repair by acting as a damage sensor and facilitator. It binds to DNA at the site of \ndamage, becomes catalytically activated, and uses NAD⁺ as a substrate to add poly (ADP-ribose) \n(PAR) chains to itself and other proteins—a process called PARylation that results in the recruitment \nof other DNA repair proteins to the damaged site. Because of the high negative charge of PAR \npolymers, extensive autoPARylation of PARP1 leads to the dissociation of PARP1 from DNA, which is \nrequired for DNA repair completion. PARP1 is often overexpressed in various cancers, including breast, \novarian, prostate, lung, and glioblastoma. This overexpression is thought to support tumor cell survival. \nSome PARP inhibitors not only block the catalytic activity of PARP1 but also trap PARP1 on DNA at \nsites of damage, preventing its release. This creates a toxic DNA-protein complex that interferes with \nDNA replication and repair, leading to cell death—particularly in cancer cells deficient in homologous \nrecombination repair (e.g., BRCA1\/2-mutant cells).\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe TR-FRET PARP1 Trapping Assay Kit is designed to detect the poly-ADP-ribosylation activity of \nPARP1 and the status of PARP1 trapping on DNA. The DNA substrate in the kit is labeled with a \nfluorophore (acceptor). A Terbium (Tb)-labeled anti-Tag2 antibody binding to Tag2-Kras serves as \nthe fluorescence donor. Activation of Tb results in fluorescence resonance energy transfer (FRET) if \nPARP1 binds to the fluorescence-labeled DNA, since the binding brings the fluorescence donor into \nclose proximity with the fluorophore acceptor. Thus, the binding status can be quantitatively \nmeasured by calculating the ratio of the emission fluorescence intensities of the acceptor (665 nm) \nand the donor (620 nm). In the presence of NAD⁺, auto-PARylation of PARP1 leads to dissociation of \nPARP1 from the DNA, resulting in a decrease in the FRET signal. Conversely, inhibition of auto-\nPARylation activity traps PARP1 on the DNA, and the FRET signal remains high.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eTR-FRET PARP1 Trapping Assay Kit \n High throughput screening of compounds that inhibit the auto-PARylation activity of PARP1 for drug \ndiscovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e7277-TAK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare PARP1 solution Thaw PARP1 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: PARP1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the PARP1 protein 500-fold (1 µL PARP1 + 499 µL assay buffer). Add 4 µl of diluted protein solution to each of positive control wells and inhibitor test wells. Add 4 µl of assay buffer to each of negative control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare the DNA substrate solution Dilute the fluorescence-labeled DNA 100-fold (1 µL DNA + 99 µL assay buffer). Add 4 µl of the diluted DNA solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare NAD+ solution Dilute the NAD+ 50-fold (1 µL NAD+ + 49 µL assay buffer). Add 5 µl of diluted NAD+ solution to each of positive control and compound test wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:100. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 99 µl assay buffer. Add 5 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e TR-FRET PARP1 Trapping Assay\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e1.9 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e Talazoparib\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eDNA Damage Response (DDR); PARP-mediated repair\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (breast; ovarian; prostate; BRCA-mutant tumors)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302212461,"sku":"72771TAK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12d-craf-cypa-inhibitor-assay-kit-bht20700014","title":"Kras G12D\/cRAF\/CYPA\/Inhibitor Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \nhuman tumors, the Ras-RAF signaling pathway is considered an important therapeutic target for cancer \n\ntreatment. However, Ras is considered undruggable since it lacks suitable binding pockets on the \nsurface. Recently, a discovery of a small molecule inhibitor blocks Ras-RAF signaling pathway by \n\nremolding Cyclophilin A (CYPA) and forming a CYPA:drug:KRAS ternary complex. This inhibitory \nstrategy provides a new method for developing drugs targeting Kras for treatment of cancers.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12D) Inhibitor assay kit is a TR-FRET based assay, which is designed to screen Kras \n\ninhibitors and determine the Kras-inhibitor binding affinity. Tag2-Kras (G12D) in this assay kit is loaded \nwith GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF has a \nTag1 at N-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a \nfluorescence donor (HTRF donor), activation of which results in fluorescence resonance energy \n\ntransfer (FRET) if Tag1-cRAF binds to the Kras, since the binding brings Terbium on the anti-Tag2 \nantibody close to the fluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status \ncan be quantitively measured by calculating the ratio of the emission fluorescence intensity of the \nacceptor (665 nm) and donor (620 nm). If an inhibitor associated with CYPA binds to the Kras and \nblocks the cRAF binding, the HTRF signal will be reduced. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12D) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-CK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000, Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare compound dilution buffer containing 2 mM DTT (CD buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare 1X Assay Buffer containing 2 mM DTT (AB buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare Kras (G12D) solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare cRAF solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate sample HTRF signal of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in CD buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in CD buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in CD buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare 1X Assay Buffer containing 2 mM DTT (AB buffer) For example, mix 500 µl of 2X Kras Binding Buffer, 496 µl of distilled water and 4 µl of 0.5 M DTT. Make only enough AB buffer as needed for the assay. Store the remaining Binding buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare Kras (G12D) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 75-fold (1µL Kras G12D + 74 µL AB buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of AB buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of CD buffer to each of negative and positive control well. -4510 or 858453-5700 Fax: 855-898-3979 3\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 400-fold (1 µL cRAF + 399 µL of AB buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in AB buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of AB buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e2.4 μM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e RMC-9805\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302179693,"sku":"5727-4123CK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12c-craf-cypa-inhibitor-assay-kit-bht20700011","title":"Kras G12C\/cRAF\/CYPA\/Inhibitor Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \nhuman tumors, the Ras-RAF signaling pathway is considered an important therapeutic target for cancer \n\ntreatment. However, Ras is considered undruggable since it lacks suitable binding pockets on the \nsurface. Recently, a discovery of a small molecule inhibitor blocks Ras-RAF signaling pathway by \n\nremolding Cyclophilin A (CYPA) and forming a CYPA:drug:KRAS ternary complex. This inhibitory \nstrategy provides a new method for developing drugs targeting Kras for treatment of cancers.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12C) Inhibitor assay kit is a TR-FRET based assay, which is designed to screen Kras \n\ninhibitors and determine the Kras-inhibitor binding affinity. Tag2-Kras (G12C) in this assay kit is loaded \nwith GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF has a \nTag1 at N-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a \nfluorescence donor (HTRF donor), activation of which results in fluorescence resonance energy \n\ntransfer (FRET) if Tag1-cRAF binds to the Kras, since the binding brings Terbium on the anti-Tag2 \nantibody close to the fluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status \ncan be quantitively measured by calculating the ratio of the emission fluorescence intensity of the \nacceptor (665 nm) and donor (620 nm). If an inhibitor associated with CYPA binds to the Kras and \nblocks the cRAF binding, the HTRF signal will be reduced. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12C) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-CK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000, Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare compound dilution buffer containing 2 mM DTT (CD buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare 1X Assay Buffer containing 2 mM DTT (AB buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare Kras (G12C) solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare cRAF solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate sample HTRF signal of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in CD buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in CD buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in CD buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare 1X Assay Buffer containing 2 mM DTT (AB buffer) For example, mix 500 µl of 2X Kras Binding Buffer, 496 µl of distilled water and 4 µl of 0.5 M DTT. Make only enough AB buffer as needed for the assay. Store the remaining Binding buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare Kras (G12C) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 110-fold (1µL Kras G12C + 109 µL AB buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of AB buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of CD buffer to each of negative and positive control well. -4510 or 858453-5700 Fax: 855-898-3979 3\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 400-fold (1 µL cRAF + 399 µL of AB buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in AB buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of AB buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e0.26 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e RMC-6291\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302310765,"sku":"5727-4122CK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12v-craf-cypa-inhibitor-assay-kit-bht20700019","title":"Kras G12V\/cRAF\/CYPA\/Inhibitor Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \nhuman tumors, the Ras-RAF signaling pathway is considered an important therapeutic target for cancer \n\ntreatment. However, Ras is considered undruggable since it lacks suitable binding pockets on the \nsurface. Recently, a discovery of a small molecule inhibitor blocks Ras-RAF signaling pathway by \n\nremolding Cyclophilin A (CYPA) and forming a CYPA:drug:KRAS ternary complex. This inhibitory \nstrategy provides a new method for developing drugs targeting Kras for treatment of cancers.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eKras (G12V)\/cRAF\/CYPA\/Inhibitor Assay Kit is a TR-FRET based assay, which is designed to screen \n\nKras inhibitors and determine the Kras-inhibitor binding affinity. Tag2-Kras (G12V) in this assay kit is \nloaded with GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF in \nthe kit has a Tag1 at N-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves \nas a fluorescence donor (HTRF donor), activation of which results in fluorescence resonance energy \n\ntransfer (FRET) if Tag1-cRAF binds to the Kras, since the binding brings Terbium on the anti-Tag2 \nantibody close to the fluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status \ncan be quantitively measured by calculating the ratio of the emission fluorescence intensity of the \nacceptor (665 nm) and donor (620 nm). If an inhibitor associated with CYPA binds to the Kras and \nblocks the cRAF binding, the HTRF signal will be reduced. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12V) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-CK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000, Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare compound dilution buffer containing 2 mM DTT (CD buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare 1X Assay Buffer containing 2 mM DTT (AB buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare Kras (G12V) solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare cRAF solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate sample HTRF signal of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in CD buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in CD buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in CD buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare 1X Assay Buffer containing 2 mM DTT (AB buffer) For example, mix 500 µl of 2X Kras Binding Buffer, 496 µl of distilled water and 4 µl of 0.5 M DTT. Make only enough AB buffer as needed for the assay. Store the remaining Binding buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare Kras (G12V) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 105-fold (1µL Kras G12V + 104 µL AB buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of AB buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of CD buffer to each of negative and positive control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare cRAF solution -4510 or 858453-5700 Fax: 855-898-3979 3 Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 400-fold (1 µL cRAF + 399 µL of AB buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in AB buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of AB buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e98 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e RMC-6236\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302343533,"sku":"5727-4128CK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"tev-protease-activity-assay-kit-bht20700001","title":"TEV Protease Activity Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eTobacco Etch Virus protease (TEV protease) is a highly sequence specific cysteine protease. It has a \nstrict 7 amino acid cleavage recognition sequence of Glu-Asn-Leu-Tyr-Phe-Gln ↓ (Gly\/Ser). The high \nspecificity makes this protease excellent for the removal of affinity-tags from purified recombinant \nproteins.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe TEV Protease Activity Assay kit is a fluorogenic-based assay to measure TEV protease activity. \nThe kit contains a TEV protease substrate that is labeled with fluorophore FAM and a quencher. \nProteolytic activity of TEV protease cleaves the substrate and releases the FAM, resulting in the \nproduction of bright fluorescence which can be measured using a fluorescence reader at ex\/em of 490 \nnM\/520 nm. TEV protease activity then can be calculated in accordance with the fluorescence intensity. \nPurified TEV protease is included in the kit as a positive control.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eMeasure TEV protease activity.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA microplate reader capable of measuring fluorescence intensity is required. Aurora Biolabs, LLC; www.aurorabiolabs.com; San Diego, CA, USA. Tel: 858-215-4510 or 858-374-6010; Tech: 858-453-5700 58-453-5700\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e190001B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e96-well microplate, black\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Dilute 1 mM 5-FAM to 20 µM with the assay buffer prepared at step A (assay buffer A).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Make 2-fold series of dilutions with the assay buffer a to get 10, 5, 2.5 1.25, 0.625, 0.3125 and 0 µM solutions.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Aliquot 50 µL of the diluted solution to each well (96-well plate).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Dilute substrate solution 25-fold with the assay buffer A.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Add 50 µl of diluted substrate to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Measure fluorescent intensity at excitation of 490 nm and emission of 520 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Use the same machine settings when measure TEV protease activity afterwards. 5-FAM Standard y t i s n e t n I e c n e c s e r o u F l 10000 8000 6000 4000 2000 0 0 2000 4000 6000 8000 10000 FAM [nM] C. Measure TEV protease positive control activity\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Thaw TEV protease protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: TEV protease protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Dilute the TEV protein 125-fold with the assay buffer A (from 1000 ng\/µL to 16 ng\/µL). Then, make a further dilution to 8, 4, 2, 0.5, 0 ng\/µL.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Add 50 µl of diluted protein solution to each well (Test amount of the protein will be 400, 200, 100, 50, 25 and 0 ng per reaction).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 11.\u003c\/strong\u003e Dilute substrate solution 25-fold with assay buffer A.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 12.\u003c\/strong\u003e Add 50 µl of diluted substrate to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 13.\u003c\/strong\u003e Incubate at room temperature for 1 hour.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 14.\u003c\/strong\u003e Measure fluorescent intensity at excitation of 490 nm and emission of 520 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 15.\u003c\/strong\u003e Plot fluorescent intensity versus protein concentration on a graph as below (subtract the average fluorescent intensity readings in the 0 ng wells from all of other wells to remove fluorescence background). TEV Activity y t i s n e t n I e c n e c s e r o u F l 7000 6000 5000 4000 3000 2000 1000 0 0 100 200 TEV [ng] 300 400 D. Measure TEV protease activity\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 16.\u003c\/strong\u003e Dilute TEV protease protein to 8, 4, 2, 0.5, 0 ng\/µL with the assay buffer A.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 17.\u003c\/strong\u003e Add 50 µl of diluted protein solution to each well (Test amount of the protein will be 400, 200, 100, 50, 25 and 0 ng per reaction). We recommend to run the reactions in duplicate.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 18.\u003c\/strong\u003e Dilute substrate solution 25-fold with assay buffer A.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 19.\u003c\/strong\u003e Add 50 µl of diluted substrate to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 20.\u003c\/strong\u003e Incubate at room temperature for 1 hour.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 21.\u003c\/strong\u003e Measure fluorescent intensity at excitation of 490 nm and emission of 520 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 22.\u003c\/strong\u003e Plot fluorescent intensity versus protein concentration on a graph as below (subtract the average fluorescent intensity readings in the 0 ng wells from all of other wells to remove fluorescence background).\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eProteolysis (affinity tag removal; protein engineering)\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eGeneral \/ Biotechnology\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"96 reactions","offer_id":53238302277997,"sku":"190001AK","price":599.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12c-nucleotide-exchange-assay-kit-bht20700012","title":"Kras G12C Nucleotide Exchange Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway plays an important role in cell proliferation and \ndifferentiation. Conversion of Kras from the inactive GDP-bound state to the active GTP-bound state \ntriggers the downstream effector and promotes cell growth. RAS genes are frequently mutated in \nvarious human tumors. These mutations block the GTPase activity of RAS and lock RAS in the GTP-\nbound state, resulting in constitutively active signals through the downstream cascades leading to \ncancer cell proliferation.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12C) nucleotide exchange assay is a TR-FRET based assay. The assay kit is designed to \ndetect the GTP binding status of Kras (G12C) in the presence of SOS1, the most-studied guanine \nnucleotide exchange factor (GEF) of Kras. The Tag2-Kras in this assay kit is recognized by a Terbium-\nlabeled anti-Tag2 antibody (HTRF donor). If Kras binds to a fluorescence-labeled GTP (HTRF \nacceptor), the donor and the acceptor will be brought in close proximity. Excitation of Terbium (340 nm) \ngenerates fluorescence resonance energy transfer (FRET) to the fluorescence-labeled GTP acceptor, \nwhich consequently fluoresces at 665 nm (figure below). Thus, GTP binding to Kras can be quantitively \nmeasured by calculation of the fluorescent ratio of 665 nm\/620 nm. The inhibitor blocking the nucleotide \nexchange will reduce the HTRF signal. \n\nAurora Biolabs LLC, San Diego, CA 92121; www.aurorabiolabs.com; \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit Kras activation for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-NK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e24 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302441837,"sku":"5727-4122NK","price":1699.0,"currency_code":"USD","in_stock":false}]},{"product_id":"wee1-binding-assay-kit-bht20700031","title":"WEE1 Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eWEE1, a nuclear kinase, belongs to WEE kinase family that negatively regulates the cell cycle \n\nvia phosphorylation of CDK1. WEE1 serves as a dual-specificity kinase which selectively \nphosphorylates both Thr14 and Tyr15 residues of both CDK1 and CDK2 to restrain their activation and \nhalt cell cycle progression in the response to DNA damage. Overexpression of WEE1 is commonly \nobserved in malignant cells and its high expression has been associated with poor rates of survival in \nvarious cancer types. Inhibition of WEE1 facilitates or even expedites mitotic progression, leading to \nan increase in genomic instability. Therefore, WEE1 is considered a potential therapeutic target for \ncancer treatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe WEE1 binding assay kit is a TR-FRET based assay, which is designed to screen compounds that \n\nbind to WEE1. If the WEE1 with N-terminal tag2 binds to a fluorescence-labelled tracer (fluorescent \n\nreceptor, emission at 665 mm), it brings the Terbium (fluorescence donor, emission at 620 mm) \n\nconjugated with anti-Tag2 antibody close to the fluorescent acceptor. Activation of the Terbium results \n\nin fluorescence resonance energy transfer (FRET), and leads to the receptor fluorescent emission at \n\n665 mm. The competitive binding of a non-fluorescence-labeled compound will reduce the receptor \n\nsignal. Thus, the compound binding status can be quantitively measured by calculating the ratio of the \n\nemission fluorescence intensity of the acceptor (665 nm) and donor (620 nm). \n\n Aurora Biolabs LLC, San Diego, CA, USA. www.aurorabiolabs.com; \n\n 1 \n\n \n \n \n \n \n \n \n \n \n \n \n\nReference\nLOt \n\nMatheson, J.C., et al., Trends Pharmacol Sci. 2016 Oct;37(10):872-881.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit WEE1 activity for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e759331-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare WEE1 solution Thaw WEE1 protein on ice. Upon first thaw, briefly spin the tube to recover all of the contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: WEE1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not store and re-use the diluted protein. Dilute the WEE1 protein 120-fold (1 µL WEE1 + 119 µL 1X assay buffer containing DTT). Add 8 µl of diluted protein solution to each positive control well and inhibitor test well. Add 8 µl of 1X DTT containing buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare fluorescence-labeled tracer and Tb-labeled anti-Tag2 antibody solution Thaw the tracer and the antibody to room temperature. Dilute the tracer 50-fold and the antibody 200-fold with 1X assay buffer containing DTT. For example, add 4 µl of the tracer and 1 µl of the anti-tag2 antibody to 200 µl of 1X DTT containing assay buffer.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Incubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eCell Cycle Regulation (DNA damage response; CDK1\/CDK2 inhibition)\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (overexpressed in malignant cells)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302376301,"sku":"759331BK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12d-nucleotide-exchange-assay-kit-bht20700015","title":"Kras G12D Nucleotide Exchange Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway plays an important role in cell proliferation and \ndifferentiation. Conversion of Kras from the inactive GDP-bound state to the active GTP-bound state \ntriggers the downstream effector and promotes cell growth. RAS genes are frequently mutated in \nvarious human tumors. These mutations block the GTPase activity of RAS and lock RAS in the GTP-\nbound state, resulting in constitutively active signals through the downstream cascades leading to \ncancer cell proliferation.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12D) nucleotide exchange assay is a TR-FRET based assay. The assay kit is designed to \ndetect the GTP binding status of Kras (G12D) in the presence of SOS1, the most-studied guanine \nnucleotide exchange factor (GEF) of Kras. The Tag2-Kras in this assay kit is recognized by a Terbium-\nlabeled anti-Tag2 antibody (HTRF donor). If Kras binds to a fluorescence-labeled GTP (HTRF \nacceptor), the donor and the acceptor will be brought in close proximity. Excitation of Terbium (340 nm) \ngenerates fluorescence resonance energy transfer (FRET) to the fluorescence-labeled GTP acceptor, \nwhich consequently fluoresces at 665 nm (figure below). Thus, GTP binding to Kras can be quantitively \nmeasured by calculation of the fluorescent ratio of 665 nm\/620 nm. The inhibitor blocking the nucleotide \nexchange will reduce the HTRF signal. \n\n Aurora Biolabs LLC, San Diego, CA 92121; www.aurorabiolabs.com; \n\nKras (G12D) Nucleotide Exchange Assay Kit\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit Kras activation for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-NK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare 1X assay buffer containing 1 mM DTT (1X DTT-containing assay buffer) For example, mix 996 µl distilled water with 1000 µl of 2X assay Buffer (Catalogue number: 5727- NK-B) and 4 µl of 0.5 M DTT. Make only enough 1X DTT-containing assay buffer as needed for the assay. Store the remaining 2X assay buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare SOS1 solution Thaw SOS1 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: SOS1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the SOS1 protein 400-fold (1 µL SOS1 + 399 µL 1X DTT-containing assay buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of 1X DTT-containing assay buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare Kras solution Kras (G12D) Nucleotide Exchange Assay Kit Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein to 440-fold (1µL Kras G12D + 439 µL 1X DTT-containing assay buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled GTP 1:40 in 1X DTT-containing assay buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled GTP + 194 µl of 1X DTT-containing assay buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e21 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302474605,"sku":"5727-4123NK","price":1699.0,"currency_code":"USD","in_stock":false}]},{"product_id":"sars-cov-2-nucleocapsid-protein-binding-kit-for-mouse-antibody-bht20700028","title":"SARS-CoV-2 Nucleocapsid Protein Binding Kit (For mouse antibody)","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eSARS-CoV-2 Nucleocapsid protein (NP) is one of the core components of SARS-CoV-2 virus. It forms \na complex with viral genomic RNA in a helical symmetrical structure and plays a key role in the process \nof virus replication and assembly. Since NP is abundantly expressed during infection, it can be used \nas an important diagnostic marker for COVID-19 and also can be used as a potential drug target or \ndeveloping vaccines.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe SARS-CoV-2 Nucleocapsid protein (NP) Binding kit is a TR-FRET based assay, that is designed \nto detect binding status of NP to the test antibody. Terbium-labeled anti-Tag5 antibody serves as \nfluorescence donor, that binds to the Tag5-NP. If a test mouse antibody binds to NP, fluorescence-\nlabeled anti-mouse antibody (fluorescence acceptor) will be brought in close proximity with the \nfluorescence donor. Excitation of Terbium (340 nm) generates fluorescence resonance energy transfer \n(FRET) to the fluorescence-labeled acceptor, which consequently fluoresces at 665 nm (figure below). \nThus, the test antibody binding to NP can be quantitively measured by calculation of the fluorescent \nratio of 665 nm\/620 nm.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of antibodies that bind to NP.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required. Aurora Biolabs LLC, San Diego, CA 92121, USA; www.aurorabiolabs.com; SARS-CoV-2 Nucleocapsid Protein Binding Kit (For mouse antibody)\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e2x Assay Buffer\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003eCustomer Test anti-NP-mouse antibody (to be tested antibody)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare SARS-CoV-2 Nucleocapsid protein Dilute SARS-CoV-2 Nucleocapsid protein (NP) 1,500-fold with 1X DTT-containing assay buffer. For example: 1 µl of NP + 1,499 µl of 1X DTT-containing assay buffer. Add 5 µl of diluted NP protein to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare Antibody solution Prepare mouse antibody with 1X DTT-containing assay buffer to the concentration to be tested. Add 5 µl of diluted antibody solution to each well except negative control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare dye solution\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Incubate the reaction at room temperature for 1 hour.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample signal. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% activity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% activity. The \npercent activity in the presence of each compound is calculated according to the following \n\nAurora Biolabs LLC, San Diego, CA 92121, USA; www. SARS-CoV-2 Nucleocapsid Protein Binding Kit \n(For mouse antibody) equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence of the \ncompound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e Nucleocapsid Protein Binding\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eViral Antigen Recognition; Vaccine Response\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eInfectious Disease (COVID-19)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302572909,"sku":"728263","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g13d-nucleotide-exchange-assay-kit-bht20700023","title":"Kras G13D Nucleotide Exchange Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway plays an important role in cell proliferation and \ndifferentiation. Conversion of Kras from the inactive GDP-bound state to the active GTP-bound state \ntriggers the downstream effector and promotes cell growth. RAS genes are frequently mutated in \nvarious human tumors. These mutations block the GTPase activity of RAS and lock RAS in the GTP-\nbound state, resulting in constitutively active signals through the downstream cascades leading to \ncancer cell proliferation.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G13D) nucleotide exchange assay is a TR-FRET based assay. The assay kit is designed to \ndetect the GTP binding status of Kras mutant (G13D). The Tag2-Kras (G13D) in this assay kit is \nrecognized by a Terbium-labeled anti-Tag2 antibody (HTRF donor). If Kras binds to a fluorescence-\nlabeled GTP (HTRF acceptor), the donor and the acceptor will be brought in close proximity. Excitation \nof Terbium (340 nm) generates fluorescence resonance energy transfer (FRET) to the fluorescence-\nlabeled GTP acceptor, which consequently fluoresces at 665 nm (figure below). Thus, GTP binding to \nKras can be quantitively measured by calculation of the fluorescent ratio of 665 nm\/620 nm. The \ninhibitor blocking the nucleotide exchange will reduce the HTRF signal.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit Kras activation for drug discovery. \n\n Aurora Biolabs LLC, San Diego, CA 92121; www.aurorabiolabs.com; \n\n 1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-NK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare 1X assay buffer containing 1 mM DTT (1X DTT-containing assay buffer) For example, mix 996 µl distilled water with 1000 µl of 2X assay Buffer (Catalogue number: 5727- NK-B) and 4 µl of 0.5 M DTT. Make only enough 1X DTT-containing assay buffer as needed for the assay. Store the remaining 2X assay buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare Kras solution Thaw Kras (G13D) protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein to 800-fold (1 µL Kras G13D + 799 µL 1X assay buffer containing DTT). Add 8 µl of diluted protein solution to the positive control and inhibitor test wells. Add 8 µl of 1X assay buffer containing DTT) to the negative control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled GTP 1:40 in 1X DTT-containing assay buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled GTP + 194 µl of 1X DTT-containing assay buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm. Negative Control Positive Control Inhibitor Test\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e Kras (G13D) Nucleotide Exchange Activity\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302736749,"sku":"5727-4133NK","price":1699.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12d-craf-binding-assay-kit-bht20700013","title":"Kras G12D–cRAF Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \n\nhuman tumors, the Ras-RAF signaling pathway is considered a potential therapeutic target for cancer \n\ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12D)-cRAF binding assay kit is a TR-FRET based assay, which is designed to detect the \n\nbinding status between Kras and cRAF. Tag2-Kras (G12D) in this assay kit is loaded with GppNHp, \n\nwhich represents the activated Kras. The Ras binding domain (RBD) of cRAF has a Tag1 at N-terminus. \n\nA Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence donor (HTRF \n\ndonor), activation of which results in fluorescence resonance energy transfer (FRET) if Tag1-cRAF \n\nbinds to the Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the fluorophore \n\non the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively measured by \n\ncalculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 \n\nnm). Blocking the Kras-cRAF binding will reduce the HTRF signal. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12D) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-BK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in Binding buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in Binding buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in Binding buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 480-fold (1 µL cRAF + 479 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of DTT containing Binding buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare Kras (G12D) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. -4510 or 858453-5700 Fax: 855-898-3979 3 Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 340-fold (1µL Kras G12D + 339 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in DTT containing Binding buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of DTT containing Binding buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e Kras (G12D)-cRAF Binding\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e7.3 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eConditions:\u003c\/em\u003e 35 nM cRAF\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302638445,"sku":"5727-4123BK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g13d-craf-cypa-inhibitor-assay-kit-bht20700022","title":"Kras G13D\/cRAF\/CYPA\/Inhibitor Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \nhuman tumors, the Ras-RAF signaling pathway is considered an important therapeutic target for cancer \n\ntreatment. However, Ras is considered undruggable since it lacks suitable binding pockets on the \nsurface. Recently, a discovery of a small molecule inhibitor blocks Ras-RAF signaling pathway by \n\nremolding Cyclophilin A (CYPA) and forming a CYPA:drug:KRAS ternary complex. This inhibitory \nstrategy provides a new method for developing drugs targeting Kras for treatment of cancers.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G13D) Inhibitor assay kit is a TR-FRET based assay, which is designed to screen Kras \n\ninhibitors and determine the Kras-inhibitor binding affinity. Tag2-Kras (G13D) in this assay kit is loaded \nwith GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF in the kit \nhas a Tag1 at N-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a \nfluorescence donor (HTRF donor), activation of which results in fluorescence resonance energy \n\ntransfer (FRET) if Tag1-cRAF binds to the Kras, since the binding brings Terbium on the anti-Tag2 \nantibody close to the fluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status \ncan be quantitively measured by calculating the ratio of the emission fluorescence intensity of the \nacceptor (665 nm) and donor (620 nm). If an inhibitor associated with CYPA binds to the Kras and \nblocks the cRAF binding, the HTRF signal will be reduced. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G13D) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-CK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000, Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare compound dilution buffer containing 2 mM DTT (CD buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare 1X Assay Buffer containing 2 mM DTT (AB buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare Kras (G13D) solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare cRAF solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate sample HTRF signal of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in CD buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in CD buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in CD buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare 1X Assay Buffer containing 2 mM DTT (AB buffer) For example, mix 500 µl of 2X Kras Binding Buffer, 496 µl of distilled water and 4 µl of 0.5 M DTT. Make only enough AB buffer as needed for the assay. Store the remaining Binding buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare Kras (G13D) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 135-fold (1µL Kras G13D + 134 µL AB buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of AB buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of CD buffer to each of negative and positive control well. -4510 or 858453-5700 Fax: 855-898-3979 3\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 400-fold (1 µL cRAF + 399 µL of AB buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in AB buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of AB buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e103 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e RMC-6236\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302671213,"sku":"5727-4133CK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"pkmyt1-binding-assay-kit-bht20700030","title":"PKMYT1 Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003ePKMYT1, a membrane-associated tyrosine- and threonine-specific cdc2-inhibitory kinase, belongs to \nWEE kinase family that plays an important role in the regulation of mitosis. PKMYT1 is involved in cell \ncycle progression and in response to DNA damages by inhibition of CDK1 activity through specific \nphosphorylation of Tyr15 and Thr14. Overexpression of PKMYT1 is observed in both solid and \nhematological tumors. Therefore, PKMYT1 is considered a potential therapeutic target for cancer \ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe PKMYT1 binding assay kit is a TR-FRET based assay, which is designed to screen compounds \nthat bind to PKMYT1. A fluorescence-labelled tracer, that can bind to PKMYT1, and the N-terminal \nGST-tagged full-length human PKMYT1 are used in this assay kit. A Terbium-labeled anti-GST \nantibody binding to the GST-PKMYT1 serves as a fluorescence donor (HTRF donor). if the \nfluorescence-labeled tracer binds to the PKMYT1, the binding brings Terbium on the anti-GST antibody \nclose to the fluorophore on the tracer (HTRF acceptor). Activation of the Terbium results in fluorescence \nresonance energy transfer (FRET). Thus, the binding status can be quantitively measured by \ncalculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 \nnm). The competitive binding of a non-fluorescence-labeled compound will reduce the FRET signal.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the PKMYT1 activity. \n\n Aurora Biolabs, LLC; www.aurorabiolabs.com; \nSan Diego, CA, USA. Tel: 858-215-4510 or 858-453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e756981-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare assay buffer containing 1 mM DTT\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare PKMYT1 solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare fluorescence-labeled tracer and Tb-labeled anti-Tag2 antibody solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate the ratio of the fluorescent intensity of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare PKMYT1 solution Thaw PKMYT1 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: PKMYT1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the PKMYT1 protein 20-fold (1 µL PKMYT1 + 19 µL 1X assay buffer containing DTT). Add 8 µl of diluted protein solution to each positive control well and inhibitor test well. Add 8 µl of 1X DTT containing buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare fluorescence-labeled tracer and Tb-labeled anti-Tag2 antibody solution Thaw the tracer and the antibody to room temperature. Dilute the tracer 100-fold and the antibody 200-fold with 1X assay buffer containing DTT. For example, add 2 µl of the tracer and 1 µl of the anti-tag2 antibody to 200 µl of 1X DTT containing assay buffer.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Incubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eCell Cycle Regulation (Mitosis entry; CDK1 inhibition)\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (solid and hematological tumors)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302605677,"sku":"756981BK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"sars-cov-2-mpro-3cl-protease-assay-kit-bht20700026","title":"SARS-CoV-2 Mpro (3CL Protease) Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eSARS-CoV-2 Mpro Assay Kit \n Mpro of Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2 Mpro, also referred to as SARS-CoV-\n\n2 Main protease, SARS-CoV-2 3CL protease) plays an essential role in viral replication by processing the \n\npolyproteins 1ab at 11 cleavage sites. Inhibiting the activity of this enzyme would block viral replication, \n\nmaking it a promising target for anti-coronaviral therapeutic agents. \n\nDescription \n\nThe Aurora SARS-CoV-2 Mpro assay kit is a homogeneous FRET-based assay for screening Mpro inhibitors. \n\nThe assay is fast and convenient, and requires just two \nsteps. In the first step, the Mpro enzyme \nis \npreincubated with the compound for 30 minutes. The \nreaction is initiated by adding substrate solution at \nthe second step. Fluorescent intensity is measured \nwith a fluorescent plate reader at the excitation \nemission \nwavelengths of 340-360 nm \nwavelengths of 460-480 nm. \n\nand \n\nItem \n2X Protease Assay Buffer \n0.5 M DTT \n5 mM Substrate \nRecombinant SARS-CoV-2 Mpro \nBlack low binding 96 well plate \n\nAmount \n20 ml \n200 µl \n10 µl \n5 µg \n1 \n\nStorage \n-20°C \n-20°C \n-80°C \n-80°C \nRT \n\nMaterials supplied \n\nCatalogue Number \n728205 \n\n728202 \n728206 \n\nMaterials Needed but not supplied \n\nA microplate reader capable of measuring fluorescence at excitation wavelengths of 340-360 nm and \nemission wavelengths of 460-480 nm. \n\nStability \n\n12 months if stored under the indicated conditions.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution. If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 5 µl to the 50 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare Mpro solution. Thaw Mpro enzyme on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Mpro enzyme is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted enzyme. Dilute the Mpro enzyme to 5 ng\/µl in 1X assay buffer. Add 20 µl of diluted enzyme solution to each of positive control well and inhibitor test well. Add 1X buffer to each of background well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add the inhibitor solution Add 5 µl of 1X assay buffer to each background well and positive control well if the inhibitor is diluted in 1X buffer. Add 5 µl of 1X assay buffer with 10% DMSO to each of background well and positive control well if the inhibitor is diluted in 1X assay buffer with 10% DMSO. Add 5 µl of diluted inhibitor solution from Step 2 to each of the inhibitor test well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Incubate at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare substrate solution During the incubation of the enzyme and the inhibitor solution, dilute the 5 mM substrate solution to 20 µM in 1X assay buffer. Make only enough solution as need for the assay. Store the remaining 5 mM Substrate solution to -80°C. Add 25 µl of diluted substrate solution to each of well, including background wells, positive control wells and the inhibitor test wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate at room temperature for 2 hours.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure the fluorescent intensity Measure the fluorescent intensity at the excitation wavelengths of 340-360 nm and the emission wavelengths of 460-480 nm\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate percentage activity of the enzyme \n\n% Activity= \n\n(Fp – Fb) – (Fi-Fb) \nFp - Fb \n\nX 100 \n\nWhere Fp refers to Fluorescent intensity of the positive control, Fb refers to Fluorescent intensity of \nbackground, and Fi refers to Fluorescent intensity of the inhibitor. Graph the percentage activity as a function of the inhibitor concentration to determine the IC50 of the test \ninhibitor. No CPD refers to no compound control \n(compound vehicle control).\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eViral Polyprotein Processing \/ Replication\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eInfectious Disease (COVID-19)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"96 reactions","offer_id":53238302507373,"sku":"728203","price":899.0,"currency_code":"USD","in_stock":false}]},{"product_id":"tr-fret-parp2-trapping-assay-kit-bht20700025","title":"TR-FRET PARP2 Trapping Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003ePARP2 (Poly (ADP-ribose) polymerase 2) is a member of the PARP family and plays a crucial role in \nDNA repair, particularly in the repair of single-strand breaks (SSBs) in DNA. It binds to DNA at the site \nof damage, becomes catalytically activated, and uses NAD⁺ as a substrate to add poly (ADP-ribose) \n(PAR) chains to itself and other proteins—a process called PARylation that results in the recruitment \nof other DNA repair proteins to the damaged site. Because of the high negative charge of PAR \npolymers, extensive autoPARylation of PARP2 leads to the dissociation of PARP2 from DNA, which is \nrequired for DNA repair completion. PARP2 is often overexpressed in various cancers, including breast, \novarian, prostate, lung, and glioblastoma. This overexpression is thought to support tumor cell survival. \nSome PARP inhibitors not only block the catalytic activity of PARP2 but also trap PARP2 on DNA at \nsites of damage, preventing its release. This creates a toxic DNA-protein complex that interferes with \nDNA replication and repair, leading to cell death, particularly in cancer cells deficient in homologous \nrecombination repair (e.g., BRCA1\/2-mutant cells).\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe TR-FRET PARP2 Trapping Assay Kit is designed to detect the poly-ADP-ribosylation activity of \nPARP2 and the status of PARP2 trapping on DNA. The DNA substrate in the kit is labeled with a \nfluorophore (acceptor). A Terbium (Tb)-labeled anti-Tag2 antibody that binds to Tag2-Kras serves as \nthe fluorescence donor. Activation of Tb results in fluorescence resonance energy transfer (FRET) if \nPARP2 binds to the fluorescence-labeled DNA, since the binding brings the fluorescence donor into \nclose proximity with the fluorophore acceptor. Thus, the binding status can be quantitatively measured \nby calculating the ratio of the emission fluorescence intensities of the acceptor (665 nm) and the donor \n(620 nm). In the presence of NAD⁺, auto-PARylation of PARP2 leads to its dissociation from DNA, \nresulting in a decrease in the FRET signal. Inhibition of auto-PARylation activity traps PARP2 on the \nDNA, and the FRET signal remains high.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the auto-PARylation activity of PARP2 for drug \ndiscovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e7277-TA-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare PARP2 solution Thaw PARP2 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: PARP2 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the PARP2 protein 500-fold (1 µL PARP2 + 499 µL assay buffer). Add 4 µl of diluted protein solution to each of positive control wells and inhibitor test wells. Add 4 µl of assay buffer to each of negative control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare the DNA substrate solution Dilute the fluorescence-labeled DNA 20-fold (1 µL DNA + 19 µL assay buffer). Add 4 µl of the diluted DNA solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare NAD+ solution Dilute the NAD+ 25-fold (1 µL NAD+ + 24 µL assay buffer). Add 5 µl of diluted NAD+ solution to each of positive control and compound test wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:100. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 99 µl assay buffer. Add 5 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e0.7 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eDNA Damage Response (DDR)\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (breast; ovarian; prostate)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302540141,"sku":"72772TAK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"tr-fret-vhl-binding-assay-kit-384-bht20700033","title":"TR-FRET VHL Binding Assay Kit (384)","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eVon Hippel–Lindau (VHL) is a member of an E3 ubiquitin ligase, a five-component complex including \nVHL, Cullin 2 (CUL2), Elongin B, Elongin C and RBX1 (RING-box protein 1). It is one of the most widely \nused E3 ligase recruiters in the design of PROTACs (Proteolysis-Targeting Chimeras) for targeted \nprotein degradation (TPD) drug discovery. VHL plays a critical role in bringing the target protein and \n\nthe ubiquitination machinery together for protein degradation via the proteasome.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe TR-FRET VHL Binding Assy kit is designed to measure the binding affinity of VHL and its ligand, \nand it includes Tag1-VHL-5C (VHL\/CUL2\/EloC\/EloB\/RBX1 complex), Terbium-labeled Anti-Tag1 \nantibody and fluorescent labeled VHL ligand VH032. The binding of VHL to the ligand brings Terbium \n(fluorescence donor) on the anti-Tag1 antibody in close proximity to the fluorophore (FL) on VH032 \n(fluorescent receptor), which results in fluorescence resonance energy transfer (FRET). Thus, the \nbinding status of VHL and VH032 can be quantitively determined using HTRF signal by calculating the \nratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 nm). If an \ncompound binds to the VHL and blocks VH032 binding, the HTRF signal will be reduced.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that bind to VHL for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required. -4510 or 858453-5700 Fax: 855-898-3979 1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e845225-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-5C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003eFluorescence-labeled VH032 (FL-VH032)\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare VHL-5C solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare FL-VH032 solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate sample HTRF signal of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare VHL-5C solution Thaw VHL-5C protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: VHL-5C protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the VHL-5C protein 40-fold (1µL VHL-5C + 39 µL assay buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of assay buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of assay buffer to each of negative and positive control wells. If the compound is diluted in 10% DMSO, add 2 µl of assay buffer containing 10% DMSO to each of\tnegative and positive control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare FL-VH032 solution Dilute FL-VH032 10-fold (1 µL FL-VH032 + 9 µL of assay buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag1 antibody 1:100 in assay buffer. For example: 1 µl of Terbium- labeled anti-Tag1 antibody + 99 µl of assay buffer. Add 10 µl of this dye mixture to each well. -4510 or 858453-5700 Fax: 855-898-3979 3\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Incubate the reaction at room temperature for 60 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e TR-FRET VHL Binding Assay\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e VH032\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eUbiquitin-Proteasome \/ Targeted Protein Degradation\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology; PROTAC Drug Discovery\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302703981,"sku":"845225","price":1299.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12c-craf-binding-assay-kit-bht20700010","title":"Kras G12C–cRAF Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \n\nhuman tumors, the Ras-RAF signaling pathway is considered a potential therapeutic target for cancer \n\ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12C)-cRAF binding assay kit is a TR-FRET based assay, which is designed to detect the \n\nbinding status between Kras and cRAF. Tag2-Kras (G12C) in this assay kit is loaded with GppNHp, \n\nwhich represents the activated Kras. The Ras binding domain (RBD) of cRAF has a Tag1 at N-terminus. \n\nA Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence donor (HTRF \n\ndonor), activation of which results in fluorescence resonance energy transfer (FRET) if Tag1-cRAF \n\nbinds to the Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the fluorophore \n\non the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively measured by \n\ncalculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 \n\nnm). Blocking the Kras-cRAF binding will reduce the HTRF signal. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12C) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-BK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in Binding buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in Binding buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in Binding buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 480-fold (1 µL cRAF + 479 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of DTT containing Binding buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare Kras (G12C) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. -4510 or 858453-5700 Fax: 855-898-3979 3 Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 340-fold (1µL Kras G12C + 339 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in DTT containing Binding buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of DTT containing Binding buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e Kras (G12D)-cRAF Binding\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e7.3 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eConditions:\u003c\/em\u003e 35 nM cRAF\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302802285,"sku":"5727-4122BK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"papain-like-plpro-protease-assay-kit-bht20700027","title":"Papain-like (PLpro) Protease Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eSARS-CoV-2 PLpro Assay Kit \n PLpro of Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2 PLpro, also referred to as SARS-CoV-\n2 Papain-like protease, Nsp3) plays an essential role in processing the polyproteins 1ab for viral replication. \nInhibition of this enzyme activity would block viral replication, making it a promising target for anti-\ncoronaviral therapeutic agents. \n\nDescription \n\nThe Aurora SARS-CoV-2 PLpro assay kit is a homogeneous assay for screening PLpro Inhibitors. \n\nThe assay is fast and convenient, and requires just \ntwo steps. In the first step, the PLpro enzyme is \npreincubated with the compound for 30 minutes. \nThe reaction is initiated by adding substrate \nsolution at the second step. Fluorescent intensity is \nmeasured with a fluorescent plate reader at the \nexcitation wavelengths of 340-360 nm and \nemission wavelengths of 460-480 nm. \n\nMaterials supplied \n\nCatalogue Number \n\nItem \n\nAmount \n\nStorage \n\n728205 \n\n728252 \n\n728256 \n\n2X Protease Assay Buffer \n\n0.5 M DTT \n\n5 mM Substrate \n\nRecombinant SARS-CoV-2 PLpro \n\nBlack low binding 96 well plate \n\n20 ml \n\n200 µl \n\n10 µl \n\n5 µg \n\n1 \n\n-20°C \n\n-20°C \n\n-80°C \n\n-80°C \n\nRT \n\nMaterials Needed but not supplied \n\nA microplate reader capable of measuring fluorescence at excitation wavelengths of 340-360 nm and \nemission wavelengths of 460-480 nm. \n\nStability \n\n12 months if stored under the indicated conditions.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution. If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 5 µl to the 50 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare PLpro solution. Thaw PLpro enzyme on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: PLpro enzyme is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted enzyme. Dilute the PLpro enzyme to 1.25 ng\/µl in 1X assay buffer. Add 20 µl of diluted enzyme solution to each of positive control well and inhibitor test well. Add 1X assay buffer to each of background well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add the inhibitor solution Add 5 µl of 1X assay buffer to each background well and positive control well if the inhibitor is diluted in 1X buffer. Add 5 µl of 1X assay buffer with 10% DMSO to each of background well and positive control well if the inhibitor is diluted in 1X buffer with 10% DMSO. Add 5 µl of diluted inhibitor solution from Step 2 to each of the inhibitor test well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Incubate at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare substrate solution During the incubation of the enzyme and the inhibitor solution, dilute the 5 mM substrate solution to 20µM in 1X assay buffer. Make only enough solution as need for the assay. Store the remaining 5 mM Substrate solution to -80°C. Add 25 µl of diluted substrate solution to each of well, including background wells, positive control wells and the inhibitor test wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate at room temperature for 2 hours.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure the fluorescent intensity\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate percentage activity of the enzyme \n\n% Activity= \n\n(Fp – Fb) – (Fi-Fb) \nFp - Fb \n\nX 100 \n\nWhere Fp refers to Fluorescent intensity of the positive control, Fb refers to Fluorescent intensity of \nbackground, and Fi refers to Fluorescent intensity of the inhibitor. Graph the percentage activity as a function of the inhibitor concentration to determine the IC50 of the test \ninhibitor. No CPD refers to no compound control \n(compound vehicle control).\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"96 reactions","offer_id":53238302835053,"sku":"728253","price":825.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12r-nucleotide-exchange-assay-kit-bht20700017","title":"Kras G12R Nucleotide Exchange Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \nsignaling pathways. The Ras signaling pathway plays an important role in cell proliferation and \ndifferentiation. Conversion of Kras from the inactive GDP-bound state to the active GTP-bound state \ntriggers the downstream effector and promotes cell growth. RAS genes are frequently mutated in \nvarious human tumors. These mutations block the GTPase activity of RAS and lock RAS in the GTP-\nbound state, resulting in constitutively active signals through the downstream cascades leading to \ncancer cell proliferation.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12R) nucleotide exchange assay is a TR-FRET based assay. The assay kit is designed to \ndetect the GTP binding status of Kras (G12R) in the presence of SOS1, the most-studied guanine \nnucleotide exchange factor (GEF) of Kras. The Tag2-Kras in this assay kit is recognized by a Terbium-\nlabeled anti-Tag2 antibody (HTRF donor). If Kras binds to a fluorescence-labeled GTP (HTRF \nacceptor), the donor and the acceptor will be brought in close proximity. Excitation of Terbium (340 nm) \ngenerates fluorescence resonance energy transfer (FRET) to the fluorescence-labeled GTP acceptor, \nwhich consequently fluoresces at 665 nm (figure below). Thus, GTP binding to Kras can be quantitively \nmeasured by calculation of the fluorescent ratio of 665 nm\/620 nm. The inhibitor blocking the nucleotide \nexchange will reduce the HTRF signal. \n\nAurora Biolabs LLC, San Diego, CA 92121, USA; www.aurorabiolabs.com; \n\nKras (G12R) Nucleotide Exchange Assay Kit\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit Kras activation for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-NK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 µL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003cli\u003e1. Prepare 1X assay buffer containing 1 mM DTT (1X DTT-containing assay buffer)\u003c\/li\u003e\n\u003cli\u003ePrepare the inhibitor compound solution\u003c\/li\u003e\n\u003cli\u003ePrepare SOS1 solution\u003c\/li\u003e\n\u003cli\u003eAdd inhibitor\u003c\/li\u003e\n\u003cli\u003ePrepare Kras solution\u003c\/li\u003e\n\u003cli\u003ePrepare dye solution\u003c\/li\u003e\n\u003cli\u003eIncubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli\u003eMeasure fluorescent intensity\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli\u003eExcitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003cli\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well.\u003c\/li\u003e\n\u003cli\u003eCalculate percentage activity\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare 1X assay buffer containing 1 mM DTT (1X DTT-containing assay buffer) For example, mix 996 µl distilled water with 1000 µl of 2X assay Buffer (Catalogue number: 5727- NK-B) and 4 µl of 0.5 M DTT. Make only enough 1X DTT-containing assay buffer as needed for the assay. Store the remaining 2X assay buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare SOS1 solution Thaw SOS1 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: SOS1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the SOS1 protein 80-fold (10 µL SOS1 + 790 µL 1X DTT-containing assay buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of 1X DTT-containing assay buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare Kras solution Kras (G12R) Nucleotide Exchange Assay Kit Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein to 380-fold (1µL Kras G12R + 379 µL 1X DTT-containing assay buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled GTP 1:40 in 1X DTT-containing assay buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled GTP + 194 µl of 1X DTT-containing assay buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e55 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302867821,"sku":"5727-4127NK","price":1699.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12v-nucleotide-exchange-assay-kit-bht20700020","title":"Kras G12V Nucleotide Exchange Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway plays an important role in cell proliferation and \ndifferentiation. Conversion of Kras from the inactive GDP-bound state to the active GTP-bound state \n\ntriggers the downstream effector and promotes cell growth. RAS genes are frequently mutated in \nvarious human tumors. These mutations block the GTPase activity of RAS and lock RAS in the GTP-\n\nbound state, resulting in constitutively active signals through the downstream cascades leading to \ncancer cell proliferation.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12V) nucleotide exchange assay is a TR-FRET based assay. The assay kit is designed to \n\ndetect the GTP binding status of wild type Kras in the presence of SOS1, the most-studied guanine \n\nnucleotide exchange factor (GEF) of Kras. The Tag2-Kras in this assay kit is recognized by a Terbium-\n\nlabeled anti-Tag2 antibody (HTRF donor). If Kras binds to a fluorescence-labeled GTP (HTRF \n\nacceptor), the donor and the acceptor will be brought in close proximity. Excitation of Terbium (340 nm) \n\ngenerates fluorescence resonance energy transfer (FRET) to the fluorescence-labeled GTP acceptor, \n\nwhich consequently fluoresces at 665 nm (figure below). Thus, GTP binding to Kras can be quantitively \n\nmeasured by calculation of the fluorescent ratio of 665 nm\/620 nm. The inhibitor blocking the nucleotide \n\nexchange will reduce the HTRF signal. \n\nAurora Biolabs LLC, San Diego, CA 92121; www.aurorabiolabs.com; \n\nKras (G12V) Nucleotide Exchange Assay Kit \n\n LOt\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit Kras activation for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-NK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare 1X aasay buffer containing 1 mM DTT (1X DTT-containing assay buffer) For example, mix 996 µl distilled water with 1000 µl of 2X assay Buffer (catalogue number: 5727- NK-B) and 4 µl of 0.5 M DTT. Make only enough 1X DTT-containing assay buffer as needed for the assay. Store the remaining 2X assay buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in 1X assay buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in 1X assay buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in 1X assay buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare SOS1 solution Thaw SOS1 protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: SOS1 protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the SOS1 protein 1,000-fold (1 µL SOS1 + 999 µL 1X DTT-containing assay buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of 1X DTT-containing assay buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare Kras solution\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled GTP 1:40 in 1X DTT-containing assay buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled GTP + 194 µl of 1X DTT-containing assay buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 20 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e23 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238303031661,"sku":"5727-4128NK","price":1699.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12v-craf-binding-assay-kit-bht20700018","title":"Kras G12V–cRAF Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \n\nhuman tumors, the Ras-RAF signaling pathway is considered a potential therapeutic target for cancer \n\ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12V)-cRAF binding assay kit is a TR-FRET based assay, which is designed to detect the \n\nbinding status between Kras and cRAF. Tag2-Kras (G12V) in this assay kit is loaded with GppNHp, \n\nwhich represents the activated Kras. The Ras binding domain (RBD) of cRAF has a Tag1 at N-terminus. \n\nA Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence donor (HTRF \n\ndonor), activation of which results in fluorescence resonance energy transfer (FRET) if the Tag1-cRAF \n\nbinds to Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the fluorophore on \n\nthe anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively measured by \n\ncalculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 \n\nnm). Blocking the Kras-cRAF binding will reduce the HTRF signal. \n\nAurora Biolabs, 10052 Mesa Ridge Court, Suite 103, San Diego, CA 92121, USA; www.aurorabiolabs.com; \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12V) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required. Amount Storage\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003eCatalog number\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in Binding buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in Binging buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in Binding buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 480-fold (1 µL cRAF + 479 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of DTT containing Binding buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare Kras (G12V) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled anti-Tag1 antibody 1:40 in DTT containing Binding buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled anti-Tag1 antibody + 194 µl of DTT containing Binding buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302900589,"sku":"5727-4128BK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g13d-craf-binding-assay-kit-bht20700021","title":"Kras G13D–cRAF Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \n\nhuman tumors, the Ras-RAF signaling pathway is considered a potential therapeutic target for cancer \n\ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G13D)-cRAF binding assay kit is a TR-FRET based assay, which is designed to detect the \n\nbinding status between Kras and cRAF. Tag2-Kras (G13D) in this assay kit is loaded with GppNHp, \n\nwhich represents the activated Kras. The Ras binding domain (RBD) of cRAF has a Tag1 at N-terminus. \n\nA Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence donor (HTRF \n\ndonor), activation of which results in fluorescence resonance energy transfer (FRET) if Tag1-cRAF \n\nbinds to the Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the fluorophore \n\non the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively measured by \n\ncalculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 \n\nnm). Blocking the Kras-cRAF binding will reduce the HTRF signal. \n\n-4510 or 858453-5700 Fax: 855-898-3979 \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G13D) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-BK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in Binding buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in Binding buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in Binding buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number of freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 400-fold (1 µL cRAF + 399 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of DTT containing Binding buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare Kras (G13D) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. -4510 or 858453-5700 Fax: 855-898-3979 3 Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 670-fold (1 µL Kras G13D + 669 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control and compound test wells. Add 4 µl of DTT containing Binding buffer to each of negative control wells.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in DTT containing Binding buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of DTT containing Binding buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eAssay:\u003c\/em\u003e Kras (G13D)-cRAF Binding\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302933357,"sku":"5727-4133BK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-wt-craf-cypa-inhibitor-assay-kit-bht20700008","title":"Kras WT\/cRAF\/CYPA\/Inhibitor Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \n\nhuman tumors, the Ras-RAF signaling pathway is considered an important therapeutic target for cancer \n\ntreatment. However, Ras is considered undruggable since it lacks suitable binding pockets on the \n\nsurface. Recently, a discovery of a small molecule inhibitor blocks Ras-RAF signaling pathway by \n\nremolding Cyclophilin A (CYPA) and forming a CYPA:drug:KRAS ternary complex. This inhibitory \n\nstrategy provides a new method for developing drugs targeting Kras for treatment of cancers.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (WT) Inhibitor assay kit is a TR-FRET based assay, which is designed to screen Kras \n\ninhibitors and determine the Kras-inhibitor binding affinity. Tag2-Kras (WT) in this assay kit is loaded \n\nwith GppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF in the kit \n\nhas a Tag1 at N-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a \n\nfluorescence donor (HTRF donor), activation of which results in fluorescence resonance energy \n\ntransfer (FRET) if Tag1-cRAF binds to the Kras, since the binding brings Terbium on the anti-Tag2 \n\nantibody close to the fluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status \n\ncan be quantitively measured by calculating the ratio of the emission fluorescence intensity of the \n\nacceptor (665 nm) and donor (620 nm). If an inhibitor associated with CYPA binds to the Kras and \n\nblocks the cRAF binding, the HTRF signal will be reduced. \n\n1\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (WT) and cRAF \nfor drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-CK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000, Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in CD buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in CD buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in CD buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare 1X Assay Buffer containing 2 mM DTT (AB buffer) For example, mix 500 µl of 2X Kras Binding Buffer, 496 µl of distilled water and 4 µl of 0.5 M DTT. Make only enough AB buffer as needed for the assay. Store the remaining Binding buffer at -20°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Prepare Kras (WT) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein 110-fold (1µL Kras WT + 109 µL AB buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of AB buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of CD buffer to each of negative and positive control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare cRAF solution 3 Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 400-fold (1 µL cRAF + 399 µL of AB buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody and fluorescence-labeled anti-Tag1 antibody 1:200 in AB buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 1 µl of fluorescence-labeled anti-Tag1 antibody + 198 µl of AB buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 10.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate sample HTRF signal of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Validation\u003c\/h4\u003e\n\u003cdiv style=\"background:#f0f7f6;border:1px solid #c8dada;border-radius:6px;padding:12px 16px;margin:8px 0\"\u003e\n\u003cstrong\u003eAssay Validation Data\u003c\/strong\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eValidated IC\u003csub\u003e50\u003c\/sub\u003e:\u003c\/em\u003e \u003cstrong\u003e101 nM\u003c\/strong\u003e\u003c\/span\u003e\u003cbr\u003e\n\u003cspan\u003e\u003cem\u003eReference Compound:\u003c\/em\u003e RMC-6236\u003c\/span\u003e\u003cbr\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBiological Pathway \/ Process\u003c\/h4\u003e\n\u003cp\u003eRAS-RAF Signaling Pathway\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eTherapeutic \/ Disease Area\u003c\/h4\u003e\n\u003cp\u003eOncology (KRAS-driven cancers)\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238302998893,"sku":"5727-4121CK","price":1999.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-g12r-craf-binding-assay-kit-bht20700016","title":"Kras G12R–cRAF Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \nproliferation, differentiation, and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \nhuman tumors, the Ras-RAF signaling pathway is considered a potential therapeutic target for cancer \n\ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (G12R)-cRAF binding assay kit is a TR-FRET based assay, which is designed to detect the \n\nbinding status between Kras and cRAF. Tag2-Kras (G12R) in this assay kit is loaded with GppNHp, \n\nwhich represents the activated Kras. The Ras binding domain (RBD) of cRAF has a Tag1 at N-terminus. \n\nA Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence donor (HTRF \n\ndonor), activation of which results in fluorescence resonance energy transfer (FRET) if Tag1-cRAF \n\nbinds to Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the fluorophore on \n\nthe anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively measured by \n\ncalculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and donor (620 \n\nnm). Blocking the Kras-cRAF binding will reduce the HTRF signal. \n\nAurora Biolabs, 10052 Mesa Ridge Court, Suite 103, San Diego, CA 92121, USA; www.aurorabiolabs.com;\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (G12R) and \ncRAF for drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-BK-B\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e5727-4122-T2P\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 µL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-80°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003eTerbium-labeled anti-Tag2 antibody\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e20 µL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-80°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in Binding buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in Binding buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in Binding buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 480-fold (1 µL cRAF + 479 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of DTT containing Binding buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each of negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare Kras (G12R) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled anti-Tag1 antibody 1:40 in DTT containing Binding buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled anti-Tag1 antibody + 194 µl of DTT containing Binding buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238303064429,"sku":"5727-4127BK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"kras-wt-craf-binding-assay-kit-bht20700007","title":"Kras WT–cRAF Binding Assay Kit","description":"\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cp\u003eKras is a member of the RAS protein family, which are a class of small GTPases involved in cell \n\nsignaling pathways. The Ras signaling pathway regulates diverse cellular processes, including cell \n\nproliferation, differentiation and survival. Conversion of Ras from the inactive GDP-bound state to the \n\nactive GTP-bound state activates the downstream effector and promotes cell growth. RAF is a key \n\ndownstream effector of RAS. Since the frequently mutated Ras genes are associated with various \n\nhuman tumors, the Ras-RAF signaling pathway is considered a potential therapeutic target for cancer \n\ntreatment.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Principle\u003c\/h4\u003e\n\u003cp\u003eThe Kras (WT, wild type)-cRAF binding assay kit is a TR-FRET based assay, which is designed to \n\ndetect the binding status between Kras and cRAF. Tag2-Kras (WT) in this assay kit is loaded with \n\nGppNHp, which represents the activated Kras. The Ras binding domain (RBD) of cRAF has a Tag1 at \n\nN-terminus. A Terbium-labeled anti-Tag2 antibody binding to the Tag2-Kras serves as a fluorescence \n\ndonor (HTRF donor), activation of which results in fluorescence resonance energy transfer (FRET) if \n\nthe Tag1-cRAF binds to Kras, since the binding brings Terbium on the anti-Tag2 antibody close to the \n\nfluorophore on the anti-Tag1 antibody (HTRF acceptor). Thus, the binding status can be quantitively \n\nmeasured by calculating the ratio of the emission fluorescence intensity of the acceptor (665 nm) and \n\ndonor (620 nm). Blocking the Kras-cRAF binding will reduce the HTRF signal. The ratio of the emission \n\nfluorescence intensity of the acceptor (665 nm) and donor (620 nm).\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eApplication\u003c\/h4\u003e\n\u003cp\u003eHigh throughput screening of compounds that inhibit the binding between activated Kras (WT) and cRAF \nfor drug discovery.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eInstrument Required\u003c\/h4\u003e\n\u003cp\u003eA HTRF® certified microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is required.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eKit Components\u003c\/h4\u003e\n\u003ctable class=\"bhc-spec-table\" style=\"width:100%;border-collapse:collapse;font-size:0.85em\"\u003e\n\u003cthead\u003e\u003ctr style=\"background:#1a5c58;color:#fff\"\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eCatalog No.\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px;text-align:left\"\u003eItem\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eAmount\u003c\/th\u003e\n\u003cth style=\"padding:4px 8px\"\u003eStorage\u003c\/th\u003e\n\u003c\/tr\u003e\u003c\/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003eCatalog number\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e25 mL\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e-20°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px\"\u003e384-well microplate, White\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003e\u003c\/td\u003e\n\u003ctd style=\"padding:4px 8px;text-align:center\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eMaterials Not Supplied\u003c\/h4\u003e\n\u003cul\u003e\n\u003cli\u003eMicroplate reader, HTRF® certified microplate reader (such as Tecan M1000 or Tecan Spark, etc.)\u003c\/li\u003e\n\u003cli\u003e0.5 M DTT\u003c\/li\u003e\n\u003cli\u003eAdjustable micro-pipettor\u003c\/li\u003e\n\u003cli\u003eSterile Tips\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eAssay Protocol\u003c\/h4\u003e\n\u003col style=\"padding-left:1.2em\"\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 1.\u003c\/strong\u003e Prepare the inhibitor compound solution If the inhibitor compound is dissolved in water, make a solution of the compound 10-fold higher than the final concentration in Binding buffer (since you will add 2 µl to the 20 µl reaction). If the inhibitor compound is dissolved in DMSO, make a 100-fold higher concentration of the compound than the highest concentration you want to test in DMSO. Then make a 10-fold dilution in Binding buffer (at this step, the compound concentration is 10-fold higher than the final concentration and the DMSO concentration is 10%). To determine an IC50 or to test lower concentrations of the compound, prepare as series of further dilutions in Binding buffer containing 10% DMSO (the final concentration of the DMSO will be 1% in all samples).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 2.\u003c\/strong\u003e Prepare cRAF solution Thaw cRAF protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted protein at -80°C. Note: cRAF protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the cRAF protein 480-fold (1 µL cRAF + 479 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each positive control well and inhibitor test well. Add 4 µl of DTT containing Binding buffer to each of negative control well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 3.\u003c\/strong\u003e Add inhibitor Add 2 µl of diluted compound solution to each inhibitor test well. Add 2 µl of inhibitor solvent solution to each negative and positive control well. Incubate at room temperature for 30 minutes (optional).\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 4.\u003c\/strong\u003e Prepare Kras (WT) solution Thaw Kras protein on ice. Upon first thaw, briefly spin tube to recover the full contents at the bottom of the tube. Make aliquots of the enzyme for single use. Store remaining undiluted enzyme at -80°C. Note: Kras protein is sensitive to freeze\/thaw cycles. Limit number freeze-thaw cycles for best results. Do not re-use the diluted protein. Dilute the Kras protein to 110-fold (1µL Kras WT + 109 µL DTT containing Binding buffer). Add 4 µl of diluted protein solution to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 5.\u003c\/strong\u003e Prepare dye solution Dilute Terbium-labeled anti-Tag2 antibody 1:200 and dilute fluorescence-labeled anti-Tag1 antibody 1:40 in DTT containing Binding buffer. For example: 1 µl of Terbium-labeled anti-Tag2 antibody + 5 µl of fluorescence-labeled anti-Tag1 antibody + 194 µl DTT containing Binding buffer. Add 10 µl of this dye mixture to each well.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 6.\u003c\/strong\u003e Incubate the reaction at room temperature for 30 minutes.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 7.\u003c\/strong\u003e Measure fluorescent intensity HTRF compatible microplate reader is needed to measure fluorescent intensity of the samples. Fluorescent intensity should be measured twice:\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 8.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 620 nm.\u003c\/li\u003e\n\u003cli style=\"margin-bottom:6px\"\u003e\n\u003cstrong\u003eStep 9.\u003c\/strong\u003e Excitation wavelength at 340 nm and emission at 665 nm.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bhc-assay-section\"\u003e\n\u003ch4\u003eData Analysis\u003c\/h4\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 1 — Calculate HTRF Signal\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003eHTRF = (Fluorescence at 665 nm \/ Fluorescence at 620 nm) × 10,000\u003c\/code\u003e\n\u003c\/div\u003e\n\u003cdiv style=\"background:#f8fbfb;border-left:3px solid #1a5c58;padding:10px 14px;margin:8px 0;border-radius:4px\"\u003e\n\u003cstrong\u003eStep 2 — Calculate % Activity\u003c\/strong\u003e\u003cbr\u003e\u003ccode style=\"font-size:0.9em\"\u003e% Activity = (S − N) \/ (P − N) × 100\u003c\/code\u003e\u003cbr\u003e\u003csmall\u003eS = sample signal  |  P = positive control (100%)  |  N = negative control (0%)\u003c\/small\u003e\n\u003c\/div\u003e\n\u003cp\u003eCalculate the HTRF signal (ratio of the fluorescent intensity at 665 mm\/620 mm) of each well. Calculate percentage activity \n\nIn the absence of the compound (positive control), the sample signal (P) is defined as 100% \nactivity. In the absence of enzyme (negative control), the sample signal (N) is defined as 0% \nactivity. The percent activity in the presence of each compound is calculated according to the \nfollowing equation: % activity = (S-N)\/(P-N) X100, where S= the sample signal in the presence \nof the compound.\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Aurora Biolabs","offers":[{"title":"384 reactions","offer_id":53238303097197,"sku":"5727-4121BK","price":1799.0,"currency_code":"USD","in_stock":false}]},{"product_id":"quantifluo-trypsin-inhibitor-assay-kit-bht15600096","title":"QuantiFluo™ Trypsin Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation of drugs and screening of potential inhibitors to trypsin proteases. Safe. Non-radioactive assay. Homogeneous and convenient. Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved. The assay uses FL485\/530nm or FP485\/530nm for signal readout. Compatible sample input includes Compounds that affect trypsin activity. Typical stated assay timing is 45 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL485\/530nm or FP485\/530nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect trypsin activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 45 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogeneous and convenient. Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved; Robust and High-throughput. Can be readily automated to assay thousands of samples per day. Robust assay with a Z’ factor of \u0026gt; 0.7. Available format information for this listing includes 100 Tests in 96-well plate (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of trypsin inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eTRYPSIN\u003c\/i\u003e(EC 3.4.21.4) is a digestive, serine protease that hydrolyzes dietary proteins in many eukaryotic and prokaryotic organisms. Trypsin predominantly cleaves peptide chains at the carboxyl side of lysine and arginine amino acids, but not before proline. BioAssay Systems’ QuantiFluo™ Trypsin Inhibitor Assay Kit uses a fluorescein isothiocyanate (FITC)-labeled synthetic substrate. The fluorescein label is highly quenched. Upon digestion by trypsin present in the sample, the substrate is cleaved into smaller peptides, which abolishes the quenching of the fluorescence label. The fluorescence or fluorescence polarization (FP) of the FITC-labeled fragments is measured at λex\/em = 485\/530 nm. Inhibition is determined by the decrease in fluorescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 485\/530 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 45 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Trypsin lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us for more details at Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify trypsin inhibitor in compounds that affect trypsin activity by FL485\/530 nm or FP485\/530 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect trypsin activity handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect trypsin activity across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests in 96-well plate (Custom bulk sizes available upon request)","offer_id":53238313517421,"sku":"DTRI-100","price":489.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/DTRIfig.jpg?v=1776668352"},{"product_id":"quantichrom-pyrophosphatase-inhibitor-assay-kit-bht15600086","title":"QuantiChrom™ Pyrophosphatase Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor high-throughput inhibitor screening and evaluation of pyrophosphatase modulators. The assay uses OD620 for signal readout. Compatible sample input includes Compounds that affect pyrophosphatase activity. Typical stated assay timing is 70 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e OD620 supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect pyrophosphatase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 70 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Fast and convenient. The procedure involves incubation of enzyme and inhibitor, addition of a single working reagent and incubation for 30 min. Room temperature assay. No 37°C incubator is needed; High-throughput. Homogenous “mix-incubate-measure” type assay. Can be readily automated on HTS liquid handling systems. Robust assay with a Z’ factor of 0.59. Can be used in 96-well, 384-well, and potentially higher density screening assays. Available format information for this listing includes 100 Tests in 96-well plate (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of pyrophosphatase inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eINORGANIC PYROPHOSPHATASE\u003c\/i\u003e(E.C.3.6.1.1) catalyzes the hydrolysis of phosphoester bonds of inorganic pyrophosphate [P\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e7\u003c\/sub\u003e\u003csup\u003e4-\u003c\/sup\u003e], thereby releasing two orthophosphate molecules. Family I PPases are essential enzymes found in all kingdoms of life and are responsible for maintaining the correct pyrophosphate equilibrium necessary to carry out nucleic acid and protein synthesis, and facilitate fatty acid beta oxidation.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eColorimetric (OD 620 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 70 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Pyrophosphatase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify pyrophosphatase inhibitor in compounds that affect pyrophosphatase by OD620 readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect pyrophosphatase handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect pyrophosphatase across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests in 96-well plate (Custom bulk sizes available upon request)","offer_id":53238314172781,"sku":"DPPI-100","price":399.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/DPPIfig.jpg?v=1776668350"},{"product_id":"quantifluo-pepsin-inhibitor-assay-kit-bht15600083","title":"QuantiFluo™ Pepsin Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eor evaluation of drugs and screening of potential inhibitors to pepsin proteases. Safe. Non-radioactive assay. Homogeneous and Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved. Robust and. The assay uses FL485\/530nm or FP485\/530nm for signal readout. Compatible sample input includes Compounds that affect pepsin activity. Typical stated assay timing is 45 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL485\/530nm or FP485\/530nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect pepsin activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 45 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogeneous and convenient. Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved; Robust and High-throughput. Can be readily automated to assay thousands of samples per day. Robust assay with a Z’ factor of \u0026gt; 0.7. Available format information for this listing includes 200 Tests in 96-well plate (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of pepsin inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003ePEPSIN\u003c\/i\u003e(EC 3.4.23.1) is a digestive, serine protease that hydrolyzes dietary proteins in many eukaryotic and prokaryotic organisms. Pepsin predominantly cleaves peptide chains at the amino side of the aromatic phenylalanine, tryptophan, and tyrosine amino acids. BioAssay Systems’ QuantiFluo™ Pepsin Inhibitor Assay Kit uses a fluorescein isothiocyanate (FITC)-labeled synthetic substrate. The fluorescein label is highly quenched. Upon digestion by pepsin present in the sample, the substrate is cleaved into smaller peptides, which abolishes the quenching of the fluorescence label. The fluorescence or fluorescence polarization (FP) of the FITC-labeled fragments is measured at λ\u003csub\u003eex\/em\u003c\/sub\u003e= 485\/530 nm. Inhibition is determined by the decrease in fluorescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 485\/530 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 45 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Pepsin lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us for more details at Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify pepsin inhibitor in compounds that affect pepsin activity by FL485\/530 nm or FP485\/530 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect pepsin activity handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect pepsin activity across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"200 Tests in 96-well plate (Custom bulk sizes available upon request)","offer_id":53238314828141,"sku":"DPEI-200","price":429.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/DPEIfig.jpg?v=1776668349"},{"product_id":"quantifluo-chymotrypsin-inhibitor-assay-kit-bht15600034","title":"QuantiFluo™ Chymotrypsin Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation of drugs and screening of potential inhibitors to chymotrypsin proteases. Safe. Non-radioactive assay. Homogeneous and convenient. Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are. The assay uses FL485\/530 nm for signal readout. Compatible sample input includes Compounds that affect chymotrypsin protease activity.. Typical stated assay timing is 45 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL485\/530 nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect chymotrypsin protease activity., which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 45 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogeneous and convenient. Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved; Robust and High-throughput. Can be readily automated to assay thousands of samples per day. Robust assay with a Z’ factor of \u0026gt;0.8. Available format information for this listing includes 100 Tests in 96-well plate (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of chymotrypsin inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eCHYMOTRYPSIN\u003c\/i\u003e(EC 3.4.21.1) is a digestive, serine protease that hydrolyzes dietary proteins in many eukaryotic and prokaryotic organisms. Chymotrypsin predominantly cleaves peptide chains at the carboxyl side of the aromatic phenylalanine, tryptophan, and tyrosine amino acids. BioAssay System’s QuantiFluo™ Chymotrypsin Inhibitor Assay Kit uses a fluorescein isothiocyanate (FITC)-labeled synthetic substrate. The fluorescein label is highly quenched. Upon digestion by chymotrypsin present in the sample, the substrate is cleaved into smaller peptides, which abolishes the quenching of the fluorescence label. The fluorescence or fluorescence polarization (FP) of the FITC-labeled fragments is measured at λ\u003csub\u003eex\/em\u003c\/sub\u003e= 485\/530 nm. Inhibition is determined by the decrease in fluorescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 485\/530 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 45 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Chymotrypsin lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us for more details at Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify chymotrypsin inhibitor in compounds that affect chymotrypsin protease by FL485\/530 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect chymotrypsin protease handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect chymotrypsin protease across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests in 96-well plate (Custom bulk sizes available upon request)","offer_id":53238315385197,"sku":"DCTI-100","price":479.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/DCTIfig.jpg?v=1776668352"},{"product_id":"quantifluo-urokinase-inhibitor-screening-assay-kit-bht15600097","title":"QuantiFluo™ Urokinase Inhibitor Screening Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation of drugs and screening potential inhibitors of urokinase. The assay uses FL380\/450nm for signal readout. Compatible sample input includes Compounds that affect urokinase activity. Typical stated assay timing is 30min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL380\/450nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect urokinase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eAnalytical range context:\u003c\/strong\u003e The supplied specifications include a stated detection limit of 0.04 U\/L for interpreting low-signal samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Fast. Assay is completed within a 30 minute reaction time; Homogeneous “mix-incubate-measure” type assay. Can be readily automated to assay thousands of samples per day. Available format information for this listing includes 100 Tests (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of urokinase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eUROKINASE PLASMINOGEN ACTIVATOR (urokinase, uPA)\u003c\/i\u003eis a key serine protease involved in the degradation of the extracellular matrix that catalyzes the conversion of plasminogen to active plasmin. It acts as a thrombolytic agent to break up blood clots and when over-expressed, has been reported to influence the growth of certain malignant tumors (breast, prostate, etc.). BioAssay Systems’ DUKI-100 Kit provides a convenient fluorimetric means to screen for potential urokinase inhibitors. In this assay, the fluorimetric substrate reacts with urokinase and an inhibitor will decrease the fluorescence at λex\/em = 380\/450 nm.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 380\/450 nm).\u003c\/p\u003e\n\n\u003ch2\u003eDetection limit and analytical sensitivity\u003c\/h2\u003e\n\u003cp\u003eReported detection limit: 0.04 U\/L.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 30min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Urokinase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify urokinase inhibitor screening in compounds that affect urokinase activity by FL380\/450 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect urokinase activity handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect urokinase activity across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests (Custom bulk sizes available upon request)","offer_id":53238317089133,"sku":"DUKI-100","price":439.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/DUKIfig.jpg?v=1776668350"},{"product_id":"quantichrom-hyaluronidase-inhibitor-screening-assay-kit-bht15600068","title":"QuantiChrom™ Hyaluronidase Inhibitor Screening Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation and high-throughput screen (HTS) of hyaluronidase modulators. The assay uses Turbidimetric (OD 600nm) for signal readout. Compatible sample input includes Compounds that affect hyaluronidase activity. Typical stated assay timing is 30 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e Turbidimetric (OD 600nm) supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect hyaluronidase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 30 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Rapid and reliable. The entire assay can be completed in 30 min.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Simple and Convenient. Simple procedure with an enzymatic reaction and addition of stop reagent. No wash or reagent transfer steps are involved; Robust and amenable to HTS. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of hyaluronidase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003cem\u003eHYALURONIDASES\u003c\/em\u003eare a family of enzymes that catalyze the degradation of the glycosaminoglycan, hyaluronic acid. Hyaluronic acid is one of the major constituents of the extracellular matrix in organisms where it contributes to both cell proliferation and migration. The role of hyaluronidases in breaking down this key factor in cell proliferation makes them a possible target for cancer treatment. One hypothesis is that increased hyaluronidase may help prevent tumor invasion by breaking down the extracellular matrix needed for tumor expansion. Conversely, decreasing hyaluronidase activity might prevent metastasis by stopping cancer cells from escaping primary tumor masses. The study to determine hyaluronidases’ exact role in cancer pathology is still ongoing.BioAssay Systems’ Hyaluronidase Inhibitor Screening Assay Kit uses a two-step turbidimetric reaction to measure hyaluronidase activity by the amount of hyaluronic acid that is hydrolyzed. A stop reagent halts the enzymatic reaction and forms turbidity with any residual hyaluronic acid in the well. The decrease in turbidity at 600 nm is directly proportional to hyaluronidase activity in the sample.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eColorimetric (OD 600 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 30 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Hyaluronidase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify hyaluronidase inhibitor screening in compounds that affect hyaluronidase activity by Turbidimetric (OD 600 nm).\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect hyaluronidase activity handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect hyaluronidase activity across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests (Custom bulk sizes available upon request)","offer_id":53238317449581,"sku":"DIHY-100","price":549.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/DIHYfig.jpg?v=1776668354"},{"product_id":"enzyfluo-farnesyltransferase-inhibitor-screening-kit-bht15600175","title":"EnzyFluo™ Farnesyltransferase Inhibitor Screening Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor high-throughput screening of FTase inhibitors and evaluation of drug modulators. The assay uses FL340\/550 nm for signal readout. Compatible sample input includes Compounds that affect farnesyltransferase activity. Typical stated assay timing is 60 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL340\/550 nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect farnesyltransferase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 60 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogeneous and convenient. “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved; High-throughput. A Z’-factor of 0.8 and higher is routinely observed in a 384-well format. Can be readily automated to assay thousands of samples per day. Available format information for this listing includes 400 Tests in 384-well plate (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of enzyfluo farnesyltransferase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eFARNESYLTRANSFERASE\u003c\/i\u003e(FTase, EC 2.5.1.58) catalyzes the transfer of a farnesyl group from farnesyl pyrophosphate to the cysteine residue of the C-terminus of target proteins. When not properly regulated, farnesylated proteins, including the Ras superfamily of small GTPases, can lead to developmental disorders and cancer. Simple, direct and high-throughput inhibitor screening assays find wide applications for oncology research. BioAssay Systems’ EIFT-400 assay kit provides a convenient fluorimetric method to screen for potential FTase inhibitors. In this assay, FTase reacts with farnesyl pyrophosphate and a dansyl-peptide substrate with measurable fluorescence at λem\/ex = 340\/550 nm. Inhibition is determined by the decrease in fluorescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 340\/550 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 60 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive FTase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us for more details at Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify enzyfluo farnesyltransferase inhibitor screening in compounds that affect farnesyltransferase by FL340\/550 nm.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect farnesyltransferase handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect farnesyltransferase across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"400 Tests in 384-well plate (Custom bulk sizes available upon request)","offer_id":53238318006637,"sku":"EIFT-400","price":409.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EIFTfig.jpg?v=1776668360"},{"product_id":"quantichrom-aldehyde-dehydrogenase-inhibitor-screening-kit-bht15600174","title":"QuantiChrom™ Aldehyde Dehydrogenase Inhibitor Screening Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation and high-throughput screen (HTS) of aldehyde dehydrogenase modulators. The assay uses OD565nm for signal readout. Compatible sample input includes Compounds that affect aldehyde dehydrogenase activity. Typical stated assay timing is 30 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e OD565nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect aldehyde dehydrogenase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 30 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Rapid and reliable. Can be completed in under an hour.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogenous and convenient. “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved; Robust and amenable to HTS: can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of aldehyde dehydrogenase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eALDEHYDE DEHYDROGENASES (ALDHs) are a superfamily of oxidoreductases that catalyze the conversion of aldehydes to carboxylic acids. ALDH is crucial in the metabolism of alcohol as alcohol dehydrogenase breaks down ethanol to acetaldehyde. Acetaldehyde, which is toxic to the body, is in turn broken down by ALDH to acetic acid. Imbalances of aldehyde dehydrogenase have been linked to both alcoholism and alcohol sensitivity in people. Inhibitors of the enzyme have been used in cases to treat alcoholism in patients. Cancer stem cell populations also display a heightened activity of ALDH, making ALDH inhibition a promising anti-cancer therapy approach. BioAssay Systems QuantiChrom™ Aldehyde Dehydrogenase Inhibitor Screening Kit is based on the enzymatic conversion of acetaldehyde to acetic acid and NADH by ALDH. The formed NADH in turn reduces a formazan reagent into a colored product the absorbance of which, measured at 565 nm, is proportional to the enzyme activity in the reaction. The percent inhibition of a test compound can be determined by comparing the activity of ALDH treated with a test compound to the activity of untreated ALDH.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eColorimetric (OD 565 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 30 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Aldehyde Dehydrogenase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify aldehyde dehydrogenase inhibitor screening in compounds that affect aldehyde dehydrogenase by OD565 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect aldehyde dehydrogenase handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect aldehyde dehydrogenase across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests","offer_id":53238318137709,"sku":"EIAL-100","price":368.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EIALfig.jpg?v=1776668361"},{"product_id":"enzyfluo-mmp-1-inhibitor-assay-kit-bht15600177","title":"EnzyFluo™ MMP-1 Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation of drugs and screening of potential inhibitors to MMP-1. Robust. Fluorescence-based assay with a longer wavelength probe (λex\/em = 490\/520 nm), minimizing test compound interference. Robust assay with a Z’ factor of \u0026gt; 0.8. The assay uses FL490\/520 nm for signal readout. Compatible sample input includes Compounds that affect MMP-1.. Typical stated assay timing is 75 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL490\/520 nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect MMP-1., which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 75 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Robust. Fluorescence-based assay with a longer wavelength probe (λ ex\/em = 490\/520 nm), minimizing test compound interference. Robust assay with a Z’ factor of \u0026gt; 0.8.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogeneous and convenient. Homogenous “mix-incubate-measure” assay. No wash and reagent transfer steps are involved; High-throughput. Can be readily automated to assay thousands of samples per day. Available format information for this listing includes 100 Tests (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of enzyfluo mmp-1 inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eMATRIX METALLOPROTEINASE-1\u003c\/i\u003eor\u003ci\u003eMMP-1\u003c\/i\u003e(EC 3.4.24.7) is a Ca\u003csup\u003e2+\u003c\/sup\u003e– and Zn\u003csup\u003e2+\u003c\/sup\u003e-dependent, interstitial collagenase that plays a role in the remodeling of the extracellular matrix of tissues. MMP-1 is up-regulated in metastatic cancer cells and in rheumatoid arthritis.BioAssay Systems’ EnzyFluo™ MMP-1 Inhibitor Assay Kit uses a quenched synthetic substrate. Upon digestion by MMP-1, the substrate is cleaved, which abolishes the quenching of the fluorescence label. The fluorescent fragment is measured at λ\u003csub\u003eex\/em\u003c\/sub\u003e= 490\/520 nm. Inhibition is determined by the decrease in fluorescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 490\/520 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 75 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive MMP-1 lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify enzyfluo mmp-1 inhibitor in compounds that affect MMP-1 by FL490\/520 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect MMP-1 handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect MMP-1 across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests (Custom bulk sizes available upon request)","offer_id":53238318268781,"sku":"EIMP1-100","price":639.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EIMP1fig.jpg?v=1776668360"},{"product_id":"enzychrom-nitric-oxide-synthase-inhibitor-screening-kit-bht15600179","title":"EnzyChrom™ Nitric Oxide Synthase Inhibitor Screening Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation and high-throughput screen (HTS) of nitric oxide synthase (NOS) modulators. The assay uses OD540nm for signal readout. Compatible sample input includes Nitric Oxide Synthase. Typical stated assay timing is 2-3 hours.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e OD540nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Nitric Oxide Synthase, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 2-3 hours helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e High-throughput. Homogenous “mix-incubate-measure” type assay. Can be readily automated on HTS liquid handling system.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Rapid and reliable. Can be completed in less than 3 hours if the assay is performed at 37°C. Available format information for this listing includes 100 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of nitric oxide synthase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eNitric oxide (NO) is a reactive radical that plays an important role in many key physiological functions. NO is the oxidation product of arginine by nitric oxide synthase (NOS) and is involved in host defense development, activation of regulatory proteins, and direct covalent interaction with functional biomolecules. Inhibition of NOS has the potential to produce diverse biological effects, particularly in the cardiovascular system. Simple, direct, and non-radioactive procedures for measuring NOS are becoming popular in research and drug discovery.\u003cbr\u003eBioAssay Systems EnzyChrom™ Nitric Oxide Synthase Inhibitor Assay Kit involves two steps: a NOS reaction step during which NO is produced followed by an NO detection step. Since the NO generated by NOS is rapidly oxidized to nitrite and nitrate, the NO production is measured following the reduction of nitrate to nitrite using an improved Griess method.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eColorimetric (OD 540 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 2-3 hours.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive NOS lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us for more details at Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify nitric oxide synthase inhibitor screening in nitric Oxide Synthase by OD540 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched nitric Oxide Synthase handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in nitric Oxide Synthase across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests","offer_id":53238318432621,"sku":"EINO-100","price":529.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EINOfig.jpg?v=1776668361"},{"product_id":"enzychrom-monoamine-oxidase-inhibitor-screening-kit-bht15600176","title":"EnzyChrom™ Monoamine Oxidase Inhibitor Screening Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation and high-throughput screen (HTS) of monoamine oxidase modulators. The assay uses FL530\/585nm for signal readout. Compatible sample input includes Compounds that affect monoamine oxidase (MAO) activity. Typical stated assay timing is 35 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL530\/585nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect monoamine oxidase (MAO) activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 35 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Homogeneous and convenient. “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Robust and amenable to HTS: can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of monoamine oxidase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003cem\u003eMONOAMINE OXIDASES\u003c\/em\u003e(MAO, EC 1.4.3.4) are a family of mitochondrial enzymes that catalyze the oxidative deamination of monoamines. Two isoforms of MAO exist, MAO-A and MAO-B, with different inhibitor selectivity and tissue distribution. MAO dysfunction is thought to be responsible for a number of neurological disorders. Unusually high or low levels of MAOs in the body have been associated with depression, schizophrenia, substance abuse, attention deficit disorder, migraines, and irregular sexual maturation. MAO inhibitors are one of the major classes of drugs prescribed for the treatment of depression, Parkinson’s, and Alzheimer’s diseases. BioAssay Systems MAO Inhibitor Screening Assay Kit provides a convenient fluorimetric means to screen for MAO enzyme inhibitors. In the assay, MAO reacts with\u003cem\u003ep\u003c\/em\u003e-tyramine, a substrate for both MAO-A and MAO-B, resulting in the formation of H\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e2\u003c\/sub\u003e, which is determined by a fluorimetric method (λ\u003csub\u003eem\/ex\u003c\/sub\u003e= 585\/530 nm). The assay is simple, sensitive, stable, and high-throughput adaptable.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 530\/585 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 35 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Monoamine Oxidase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify monoamine oxidase inhibitor screening in compounds that affect monoamine oxidase (MAO) by FL530\/585 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect monoamine oxidase (MAO) handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect monoamine oxidase (MAO) across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests (Custom bulk sizes available upon request)","offer_id":53238318629229,"sku":"EIMO-100","price":459.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EIMOfig.jpg?v=1776668360"},{"product_id":"enzyfluo-mmp-9-inhibitor-assay-kit-bht15600178","title":"EnzyFluo™ MMP-9 Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation of drugs and screening of potential inhibitors to MMP-9. Robust. Fluorescence-based assay with a longer wavelength probe (λex\/em = 490\/520 nm), minimizing test compound interference. Robust assay with a Z’ factor of \u0026gt; 0.8. The assay uses FL490\/520nm for signal readout. Compatible sample input includes Compounds that affect MMP-9 activity.. Typical stated assay timing is 75 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FL490\/520nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect MMP-9 activity., which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 75 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Robust. Fluorescence-based assay with a longer wavelength probe (λ ex\/em = 490\/520 nm), minimizing test compound interference. Robust assay with a Z’ factor of \u0026gt; 0.8.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Homogeneous and convenient. Homogenous “mix-incubate-measure” assay. No wash and reagent transfer steps are involved; High-throughput. Can be readily automated to assay thousands of samples per day. Available format information for this listing includes 100 Tests in 96-well plate.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of enzyfluo mmp-9 inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eMATRIX METALLOPROTEINASE-9\u003c\/i\u003eor\u003ci\u003eMMP-9\u003c\/i\u003e(EC 3.4.24.35) is a Ca\u003csup\u003e2+\u003c\/sup\u003e-and Zn\u003csup\u003e2+\u003c\/sup\u003e-dependent, interstitial gelatinase that plays a role in the remodeling of the extracellular matrix of tissues. MMP-9 plays a central role in processes diverse as wound healing, angiogenesis, and bone development. In addition to its role in normal growth and development, MMP-9 is up-regulated in many diseases, including metastatic cancer, rheumatoid arthritis, and asthma. BioAssay Systems’ EnzyFluo™ MMP-9 Inhibitor Assay Kit uses a quenched synthetic substrate. Upon digestion by MMP-9, the substrate is cleaved, which abolishes the quenching of the fluorescence label.  The fluorescent fragment is measured at λ\u003csub\u003eex\/em\u003c\/sub\u003e= 490\/520 nm. Inhibition is determined by the decrease in fluorescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 490\/520 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 75 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive MMP-9 lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify enzyfluo mmp-9 inhibitor in compounds that affect MMP-9 activity by FL490\/520 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect MMP-9 activity handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect MMP-9 activity across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests in 96-well plate","offer_id":53238319251821,"sku":"EIMP9-100","price":639.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EIMP9fig.jpg?v=1776668358"},{"product_id":"parallel-artificial-membrane-permeability-assay-pampa-kit-bht15600226","title":"Parallel Artificial Membrane Permeability Assay (PAMPA) Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor quantitative determination of gastrointestinal (GI) membrane permeability of test compounds. The assay uses PAMPA for signal readout. Compatible sample input includes Chemical compounds. Typical stated assay timing is Assay takes 20 hr, hands-on time 1 hr.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e PAMPA supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Chemical compounds, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of Assay takes 20 hr, hands-on time 1 hr helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Convenient. Includes all necessary equipment to run a PAMPA plate.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Simple and low-cost. Procedure is easy to follow and more affordable than cell-based permeability assays; High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 96 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of parallel artificial membrane permeability assay (pampa) within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eMEMBRANE PERMEABILITY is an important characteristic to determine for evaluating compounds as potential drug candidates. Drugs often need to cross cell membranes in order to reach their target of action and this makes a compound’s ability to passively cross these membranes an important characteristic to evaluate. Permeability can be evaluated by cell-based methods; however, these methods are often expensive and time consuming. Parallel Artificial Permeability Assays (PAMPA) offer researchers a quick, inexpensive method of evaluating the gastrointestinal (GI) permeability of test compounds. BioAssay Systems’ PAMPA Kit provides all the necessary components to run a Parallel Artificial Permeability Assay.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003ePAMPA.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: Assay takes 20 hr, hands-on time 1 hr.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eShort assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify parallel artificial membrane permeability assay (pampa) in chemical compounds by PAMPA readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched chemical compounds handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in chemical compounds across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"96 Tests","offer_id":53238319415661,"sku":"PAMPA-096","price":509.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/PAMPAfig.jpg?v=1776668362"},{"product_id":"parallel-artificial-membrane-permeability-assay-bbb-kit-bht15600227","title":"Parallel Artificial Membrane Permeability Assay-BBB Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor quantitative determination of blood brain barrier (BBB) membrane permeability of test compounds. The assay uses PAMPA for signal readout. Compatible sample input includes Chemical compounds. Typical stated assay timing is Assay takes 20 hr, hands-on time 1 hr.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e PAMPA supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Chemical compounds, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of Assay takes 20 hr, hands-on time 1 hr helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Convenient. Includes all necessary equipment to run a PAMPA plate.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Simple and low-cost. Procedure is easy to follow and more affordable than cell-based permeability assays; High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 96 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of parallel artificial membrane permeability assay-bbb within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eMEMBRANE PERMEABILITY\u003c\/i\u003eis an important characteristic to determine for evaluating compounds as potential drug candidates. Drugs often need to cross cell membranes in order to reach their target of action and this makes a compound’s ability to passively cross these membranes an important characteristic to evaluate. The Blood Brain Barrier (BBB) is made of brain endothelial cells with tight junctions. Rapid and early screening of compounds for BBB penetration is highly desirable for drug discovery. Permeability can be evaluated by cell-based methods; however, these methods are often expensive and time-consuming. Parallel Artificial Permeability Assays (PAMPA) offer researchers a quick, inexpensive method of evaluating the permeability of test compounds. Our PMBBB-096 kit is designed to aid in evaluating BBB permeability. BioAssay Systems’ PMBBB Kit provides all the necessary components to run a Parallel Artificial Permeability Assay for Blood Brain Barrier studies.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003ePAMPA.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: Assay takes 20 hr, hands-on time 1 hr.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eShort assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify parallel artificial membrane permeability assay-bbb in chemical compounds by PAMPA readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched chemical compounds handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in chemical compounds across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"96 Tests","offer_id":53238319448429,"sku":"PMBBB-096","price":519.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/PMBBBfig.jpg?v=1776668362"},{"product_id":"quantichrom-acetylcholinesterase-inhibitor-assay-kit-bht15600221","title":"QuantiChrom™ Acetylcholinesterase Inhibitor Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation and high-throughput screen (HTS) of acetylcholinesterase modulators. The assay uses OD412nm for signal readout. Compatible sample input includes Compounds that affect acetylcholinesterase activity. Typical stated assay timing is 30 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e OD412nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect acetylcholinesterase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 30 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Fast and sensitive. Linear detection range from 10 to 600 U\/L for a 10-minute reaction at room temperature.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests (Custom bulk sizes available upon request).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of acetylcholinesterase inhibitor within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003cem\u003eACETYLCHOLINESTERASE\u003c\/em\u003e(EC 3.1.1.7, AChE), also known as RBC cholinesterase, is found primarily in the blood and neural synapses. AChE catalyzes the hydrolysis of the neurotransmitter acetylcholine into choline and acetic acid, a reaction necessary to allow a cholinergic neuron to return to its resting state after activation. Inhibition of the enzyme leads to acetylcholine accumulation, hyperstimulation of nicotinic and muscarinic receptors, and disrupted neurotransmission. AChE inhibition is an important target for the management of Alzheimer’s disease. In addition to Alzheimer’s disease, AChE inhibitors have been useful in the diagnosis or treatment of diseases such as glaucoma, myasthenia gravis, bladder distention, and more. BioAssay Systems QuantiChrom™ Acetylcholinesterase Inhibitor Assay is based on an improved Ellman method, in which thiocholine produced by the action of acetylcholinesterase forms a yellow color with 5,5′-dithiobis(2-nitrobenzoic acid). The intensity of the product color, measured at 412nm, is proportionate to the enzyme activity in the sample.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eColorimetric (OD 412 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 30 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive Acetylcholinesterase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us by Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify acetylcholinesterase inhibitor in compounds that affect acetylcholinesterase by OD412 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect acetylcholinesterase handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect acetylcholinesterase across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests (Custom bulk sizes available upon request)","offer_id":53238319677805,"sku":"IACE-100","price":449.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/IACEfig.jpg?v=1776668363"},{"product_id":"parallel-artificial-membrane-permeability-assay-skin-kit-bht15600228","title":"Parallel Artificial Membrane Permeability Assay-Skin Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor quantitative determination of skin membrane permeability of test compounds. The assay uses PAMPA for signal readout. Compatible sample input includes Chemical Compounds. Typical stated assay timing is Assay takes 20 hrs, hands-on time 1 hr.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e PAMPA supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Chemical Compounds, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of Assay takes 20 hrs, hands-on time 1 hr helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Convenient. Includes all necessary equipment to run a PAMPA plate.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Simple and low-cost. Procedure is easy to follow and more affordable than cell-based permeability assays; High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 96 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of parallel artificial membrane permeability assay-skin within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eMEMBRANE PERMEABILITY\u003c\/i\u003eis an important characteristic to determine for evaluating compounds as potential drug candidates. Drugs often need to cross cell membranes in order to reach their target of action and this makes a compound’s ability to passively cross these membranes an important characteristic to evaluate. The skin, and in particular, the stratum corneum is a complex barrier, which can be mimicked. Rapid and early screening of compounds for skin penetration is highly desirable for drug discovery. Permeability can be evaluated by cell-based methods; however, these methods are often expensive and time-consuming. Parallel Artificial Permeability Assays (PAMPA) offer researchers a quick, inexpensive method of evaluating the permeability of test compounds. Our PMSKN-096 kit is designed to aid in evaluating skin permeability. BioAssay Systems’ PMSKN Kit provides all the necessary components to run a Parallel Artificial Permeability Assay for skin permeability studies.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003ePAMPA.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: Assay takes 20 hrs, hands-on time 1 hr.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eShort assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify parallel artificial membrane permeability assay-skin in chemical Compounds by PAMPA readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched chemical Compounds handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in chemical Compounds across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"96 Tests","offer_id":53238320136557,"sku":"PMSKN-096","price":519.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/PMSKNfig.jpg?v=1776668364"},{"product_id":"quantifluo-alpha-amylase-inhibitor-screening-kit-bht15600222","title":"QuantiFluo™ α-Amylase Inhibitor Screening Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor evaluation of drugs and screening potential inhibitors of α-amylase. Safe: Non-radioactive assay. Fast and convenient: Homogeneous “mix-incubate-measure” type assay. Can be completed in under an hour at room temperature. Robust and. The assay uses FP485\/520 nm for signal readout. Compatible sample input includes Compounds that affect α-amylase activity. Typical stated assay timing is 40 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e FP485\/520 nm supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Compounds that affect α-amylase activity, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of 40 min helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Safe. Non-radioactive assay.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Fast and convenient. Homogeneous “mix-incubate-measure” type assay. Can be completed in under an hour at room temperature; Robust and High-throughput. The FP assay greatly reduces background matrix interferences. A Z’-factor of \u0026gt;0.90 was observed in a 384-well format. Can be readily automated to assay thousands of samples per day. Available format information for this listing includes 400 Tests in 384-well plate (200 Tests in 96-well plates).\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of α-amylase inhibitor screening within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003e\u003ci\u003eAMYLASE\u003c\/i\u003ebelongs to the family of glycoside hydrolase enzymes that break down starch into glucose molecules by acting on α-1,4-glycosidic bonds. The α-amylases (EC 3.2.1.1) cleave at random locations on the starch chain, ultimately yielding maltotriose and maltose, glucose, and “limit dextrin” from amylose and amylopectin. In mammals, α-amylase is a major digestive enzyme. Increased enzyme levels in humans are associated with salivary trauma, mumps due to inflammation of the salivary glands, pancreatitis, and renal failure.Simple, direct, and automation-ready procedures for measuring α-amylase inhibition are highly desirable in Research and Drug Discovery. BioAssay Systems’ α-Amylase Inhibitor Screening Kit utilizes fluorescence polarization (FP), a highly reliable and robust technique that significantly reduces background matrix interferences, enabling the effective identification of potential α-amylase inhibitors. In this assay, α-amylase cleaves a fluorescent substrate. The decrease in FP is directly proportional to the α-amylase activity in the sample. Inhibition is therefore determined by the increase in FP (λex\/em = 485\/520 nm).\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eFluorescent (FL 485\/520 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 40 min.\u003c\/p\u003e\n\n\u003ch2\u003eScreening services\u003c\/h2\u003e\n\u003cp\u003eBioAssay Systems offers comprehensive α-Amylase lead discovery services, including compound library screening, inhibitor profiling, and high-throughput assays to identify potential drug candidates. We provide detailed analyses of compound activity, potency, and selectivity, supporting hit-to-lead and lead optimization efforts. Please contact us for more details at Service .\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify α-amylase inhibitor screening in compounds that affect α-amylase activity by FP485\/520 nm readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched compounds that affect α-amylase activity handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in compounds that affect α-amylase activity across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"400 Tests in 384-well plate (200 Tests in 96-well plates)","offer_id":53238320169325,"sku":"IAMY-400","price":489.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/IAMYfig.jpg?v=1776668362"},{"product_id":"superlight-luciferase-reporter-gene-assay-kit-bht15600257","title":"SuperLight™ Luciferase Reporter Gene Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eBright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. The assay uses Luminescence for signal readout. Compatible sample input includes Cells etc. Typical stated assay timing is 2 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e Luminescence supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Cells etc, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eAnalytical range context:\u003c\/strong\u003e The supplied specifications include a stated detection limit of 2 fg luciferase for interpreting low-signal samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR\/LeadSeeker). Assay reagents compatible with all liquid handling systems; Fast and convenient. Homogeneous “mix-and-measure” assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Available format information for this listing includes 500 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of superlight luciferase reporter gene within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eThe SuperLight™ Luciferase Reporter Gene Assay is based on the quantitation of luciferase expression in mammalian, yeast or E. coil cells, using luciferin and ATP as substrates. The reaction results in light production which can be conveniently measured on a luminometer. This bioluminescent reporter gene assay is extremely sensitive and is especially suitable for quantifying luciferase expression in recombinant cells. This ultra-sensitive, homogeneous cell-based assay only requires adding a single reagent to the cells and measuring the light intensity after a short incubation step (2 minutes). Assays can be performed in tubes, cuvettes or multi-well plates. All kit components are compatible with culture media and with all liquid handling systems. With an extended luminescence emission kinetics (half-life 40 min), the SuperLight™ luciferase assays are especially suitable for high-throughput screening in 96-well, 384-well and 1536-well plates. In addition, the reagent provided in the kits has been formulated for maximum sensitivity, reproducibility and long shelf-life. Applications for this kit include gene regulation studies and high-throughput screening of gene modulators\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eLuminescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection limit and analytical sensitivity\u003c\/h2\u003e\n\u003cp\u003eReported detection limit: 2 fg luciferase.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 2 min.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify reporter or luminescence output in cells by Luminescence readout.\u003c\/li\u003e\n  \u003cli\u003eCompare perturbation-dependent signal changes across matched sample groups.\u003c\/li\u003e\n  \u003cli\u003eMonitor reporter time-courses following stimulation, inhibition, or media changes.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"500 Tests","offer_id":53238320300397,"sku":"SLLU-500","price":339.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/SLLUfig_dca6bb52-407a-4373-8b1d-0f65a37a7de2.jpg?v=1776668364"},{"product_id":"superlight-luciferase-reporter-gene-assay-kit-bht15600255","title":"SuperLight™ Luciferase Reporter Gene Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eBright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. The assay uses Luminescence for signal readout. Compatible sample input includes Cells etc. Typical stated assay timing is 2 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e Luminescence supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Cells etc, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eAnalytical range context:\u003c\/strong\u003e The supplied specifications include a stated detection limit of 2 fg luciferase for interpreting low-signal samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR\/LeadSeeker). Assay reagents compatible with all liquid handling systems; Fast and convenient. Homogeneous “mix-and-measure” assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Available format information for this listing includes 1000 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of superlight luciferase reporter gene within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eThe SuperLight™ Luciferase Reporter Gene Assay is based on the quantitation of luciferase expression in mammalian, yeast or E. coil cells, using luciferin and ATP as substrates. The reaction results in light production which can be conveniently measured on a luminometer. This bioluminescent reporter gene assay is extremely sensitive and is especially suitable for quantifying luciferase expression in recombinant cells. This ultra-sensitive, homogeneous cell-based assay only requires adding a single reagent to the cells and measuring the light intensity after a short incubation step (2 minutes). Assays can be performed in tubes, cuvettes or multi-well plates. All kit components are compatible with culture media and with all liquid handling systems. With an extended luminescence emission kinetics (half-life 40 min), the SuperLight™ luciferase assays are especially suitable for high-throughput screening in 96-well, 384-well and 1536-well plates. In addition, the reagent provided in the kits has been formulated for maximum sensitivity, reproducibility and long shelf-life. Applications for this kit include gene regulation studies and high-throughput screening of gene modulator\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eLuminescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection limit and analytical sensitivity\u003c\/h2\u003e\n\u003cp\u003eReported detection limit: 2 fg luciferase.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 2 min.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eThe description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify reporter or luminescence output in cells by Luminescence readout.\u003c\/li\u003e\n  \u003cli\u003eCompare perturbation-dependent signal changes across matched sample groups.\u003c\/li\u003e\n  \u003cli\u003eMonitor reporter time-courses following stimulation, inhibition, or media changes.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"1000 Tests","offer_id":53238320726381,"sku":"SLLU-01K","price":509.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/SLLUfig.jpg?v=1776668363"},{"product_id":"superlight-dual-luciferase-reporter-gene-assay-kit-bht15600254","title":"SuperLight™ Dual Luciferase Reporter Gene Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eBioluminescent reagent system for rapid quantitation of firelfy and Ranilla luciferase reporter gene expression in transfected cells. The assay uses Luminescence for signal readout. Compatible sample input includes Cells etc. Typical stated assay timing is 20 min.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e Luminescence supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Cells etc, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eAnalytical range context:\u003c\/strong\u003e The supplied specifications include a stated detection limit of 2 fg luciferase for interpreting low-signal samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e High sensitivity and wide detection range. Detection of as little as 2 fg luciferase.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, and measured with any luminometer. Can be readily automated on HTS liquid handling systems; Fast and convenient. Three-step assay allows the detection of dual luciferase levels within 20 minutes. Available format information for this listing includes 100 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of superlight dual luciferase reporter gene within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eThe accuracy of reporter assays can be improved by utilizing a dual reporter system. One of the reporter genes is correlated with the promoter of interest and is used to assess the effects of specific experimental conditions, while the second reporter is used as a control and serves as a baseline response. The SuperLight™ Dual-Luciferase Reporter Gene Assay allows for the sequential measurement of the activity of two different luciferases, firefly (FFL) and Renilla (RL), in a single sample. The firefly luciferase luminescence is measured first by the addition of the FFL Reagent. Next, the RL Reagent is added to the same well. The RL Reagent simultaneously quenches the firefly luciferase luminescence and initiates the Renilla luciferase reaction. The light production of both reactions can be conveniently measured on a luminometer. This bioluminescent dual reporter gene assay is extremely sensitive and is especially suitable for quantifying dual luciferase expression in recombinant cells or in cell-free transcription\/translation reactions. Assays can be performed in tubes, cuvettes, or multi-well plates.\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eLuminescence.\u003c\/p\u003e\n\n\u003ch2\u003eDetection limit and analytical sensitivity\u003c\/h2\u003e\n\u003cp\u003eReported detection limit: 2 fg luciferase.\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: 20 min.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eShort assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify reporter or luminescence output in cells by Luminescence readout.\u003c\/li\u003e\n  \u003cli\u003eCompare perturbation-dependent signal changes across matched sample groups.\u003c\/li\u003e\n  \u003cli\u003eMonitor reporter time-courses following stimulation, inhibition, or media changes.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests","offer_id":53238320857453,"sku":"SLDL-100","price":229.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/SLDLfig.jpg?v=1776668363"}],"url":"https:\/\/www.ebiohippo.com\/collections\/rs-ic50-and-dose-response.oembed?page=2","provider":"BioHippo","version":"1.0","type":"link"}