{"product_id":"human-topoisomerase-i-dna-relaxation-assay-kit-bht16300011","title":"Human Topoisomerase I DNA Relaxation Assay Kit","description":"\u003cstyle\u003e.bhp-desc{font-size:16px;color:#1a1a1a;line-height:1.7}.bhp-desc h2{font-size:18px;font-weight:700;color:#003366;margin:24px 0 10px;padding-bottom:6px;border-bottom:2px solid #003366}.bhp-desc p{margin:0 0 12px}.bhp-desc .bhp-spec-table{width:100%;border-collapse:collapse;margin:12px 0 18px}.bhp-desc .bhp-spec-table th{text-align:left;padding:8px 12px;background:#e8f0fb;color:#003366;font-weight:600;vertical-align:top;border:1px solid #ccd9f0}.bhp-desc .bhp-spec-table td{padding:8px 12px;vertical-align:top;border:1px solid #ccd9f0}.bhp-desc .bhp-warning-box{background:#fff8e6;border-left:4px solid #e6a817;padding:10px 14px;margin:10px 0 14px;border-radius:0 4px 4px 0}.bhp-desc .bhp-cat-pill{display:inline-block;background:#003366;color:#fff;border-radius:20px;padding:3px 12px;font-size:13px;margin-bottom:10px}.bhp-desc ol,.bhp-desc ul{margin:0 0 12px 20px}\u003c\/style\u003e\u003cdiv class=\"bhp-desc\"\u003e\n\u003cspan class=\"bhp-cat-pill\"\u003eDrug Target Assay Kit\u003c\/span\u003e\u003cp\u003eHuman topoisomerase I relaxes supercoiled circular DNA. In this kit, relaxed DNA gives much weaker fluorescence with dye H19 (with magnesium present) than supercoiled DNA does, so the signal drops as the enzyme works.\u003c\/p\u003e\n\u003ch2\u003eFormats and Pack Sizes\u003c\/h2\u003e\n\u003cp\u003eHRA100K has no enzyme (human topo I is bought separately). HRA100KE adds 10 µl of 500 U\/µl human topoisomerase I for 100 assays, and HRA1000KE is the larger enzyme-included kit for 1000 assays in 96-well format (2000 in 384-well).\u003c\/p\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eKit (enzyme not included), 100 assays (HRA100K)\u003c\/th\u003e\n\u003ctd\u003e100 assays (96-well) or 200 assays (384-well)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eKit Plus (enzyme included), 100 assays (HRA100KE)\u003c\/th\u003e\n\u003ctd\u003e100 assays (96-well) or 200 assays (384-well)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eKit Plus (enzyme included), 1000 assays (HRA1000KE)\u003c\/th\u003e\n\u003ctd\u003e1000 assays (96-well) or 2000 assays (384-well)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003ch2\u003eKit Components\u003c\/h2\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eComponent\u003c\/th\u003e\n\u003cth\u003eKit (enzyme not included), 100 assays (HRA100K)\u003c\/th\u003e\n\u003cth\u003eKit Plus (enzyme included), 100 assays (HRA100KE)\u003c\/th\u003e\n\u003cth\u003eKit Plus (enzyme included), 1000 assays (HRA1000KE)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Buffer HT1\u003c\/td\u003e\n\u003ctd\u003e480 µl\u003c\/td\u003e\n\u003ctd\u003e480 µl\u003c\/td\u003e\n\u003ctd\u003e4800 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Supercoiled plasmid DNA\u003c\/td\u003e\n\u003ctd\u003e405 µl\u003c\/td\u003e\n\u003ctd\u003e405 µl\u003c\/td\u003e\n\u003ctd\u003e4050 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e1500 x Dye H19\u003c\/td\u003e\n\u003ctd\u003e20 µl\u003c\/td\u003e\n\u003ctd\u003e20 µl\u003c\/td\u003e\n\u003ctd\u003e170 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x H19 dilution buffer\u003c\/td\u003e\n\u003ctd\u003e3000 µl\u003c\/td\u003e\n\u003ctd\u003e3000 µl\u003c\/td\u003e\n\u003ctd\u003e30 ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e1000 x human topoisomerase I (500 U\/µl)\u003c\/td\u003e\n\u003ctd\u003eNot included\u003c\/td\u003e\n\u003ctd\u003e10 µl\u003c\/td\u003e\n\u003ctd\u003e40 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eFinal reaction concentrations: 25 mM Tris-HCl, pH 7.4, 58 mM KCl, 0.25 mM DTT, 5 mM MgCl2, 0.25 mM EDTA, 15 µg\/ml BSA, 25 µg\/ml supercoiled plasmid DNA and 0.1 U\/µl topoisomerase I.\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eWith magnesium present, relaxed DNA suppresses the fluorescence of dye H19 much more than supercoiled DNA does. As topo I converts supercoiled DNA into its relaxed form, the fluorescence signal at 535 nm (excitation 485 nm) goes down.\u003c\/p\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Target\u003c\/th\u003e\n\u003ctd\u003eHuman topo I DNA relaxation\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eTarget Type\u003c\/th\u003e\n\u003ctd\u003eTopoisomerase\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eOrganism\u003c\/th\u003e\n\u003ctd\u003eHuman\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Type\u003c\/th\u003e\n\u003ctd\u003eEnzyme Activity Assay\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eDetection Method\u003c\/th\u003e\n\u003ctd\u003eFluorescence\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eDetection Wavelength\u003c\/th\u003e\n\u003ctd\u003eEx 485 nm \/ Em 535 nm\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003ePlate Format\u003c\/th\u003e\n\u003ctd\u003e96-well or 384-well plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eInstrument Required\u003c\/th\u003e\n\u003ctd\u003eFluorescence microplate reader\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Time\u003c\/th\u003e\n\u003ctd\u003eAbout 65 min (60 min reaction + 5 min dye incubation)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSizes\u003c\/th\u003e\n\u003ctd\u003e100 assays (96-well) or 200 assays (384-well), 1000 assays (96-well) or 2000 assays (384-well)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eStorage\u003c\/th\u003e\n\u003ctd\u003ePlease contact us to confirm storage conditions.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eShipping\u003c\/th\u003e\n\u003ctd\u003eShipped frozen.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003ch2\u003eProtocol\u003c\/h2\u003e\n\u003cp\u003eThe manufacturer's protocol describes the procedure and reaction volumes.\u003c\/p\u003e\n\u003cul\u003e\u003cli\u003eHuman Topo I DNA Relaxation Assay (PDF, see Documents): Covers catalog # HRA100K, HRA100KE, HRA1000KE.\u003c\/li\u003e\u003c\/ul\u003e\n\u003ch2\u003eSafety \u0026amp; Handling\u003c\/h2\u003e\n\u003cdiv class=\"bhp-warning-box\"\u003eFor Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The Human Topoisomerase I DNA Relaxation Assay Kit is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Please contact us to confirm storage conditions.\u003c\/div\u003e\n\u003c\/div\u003e","brand":"ProFoldin","offers":[{"title":"Kit (enzyme not included) \/ 100 assays","offer_id":67794876465517,"sku":"HRA100K","price":825.0,"currency_code":"USD","in_stock":true},{"title":"Kit Plus (enzyme included) \/ 100 assays","offer_id":67794883510637,"sku":"HRA100KE","price":1057.0,"currency_code":"USD","in_stock":true},{"title":"Kit Plus (enzyme included) \/ 1000 assays","offer_id":67794883543405,"sku":"HRA1000KE","price":5496.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/hTopo_I_HTS_assay_with_IC50_copy.jpg?v=1792791969","url":"https:\/\/www.ebiohippo.com\/products\/human-topoisomerase-i-dna-relaxation-assay-kit-bht16300011","provider":"BioHippo","version":"1.0","type":"link"}