{"product_id":"dna-topoisomerase-i-assay-kit-bht16300061","title":"DNA Topoisomerase I 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\u003eDNA relaxation is the change of supercoiled circular DNA into relaxed DNA, carried out by DNA topoisomerases such as bacterial topoisomerase I.\u003c\/p\u003e\n\u003ch2\u003eFormats and Pack Sizes\u003c\/h2\u003e\n\u003cp\u003eDRA100K supplies the assay reagents without enzyme, while DRA100KE adds 42 µl of 100 x E. coli topoisomerase I. The H19 dye is also packed differently (8 µl of 4500 x in DRA100K vs 20 µl of 1500 x in DRA100KE).\u003c\/p\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eKit (enzyme not included), 100 assays (DRA100K)\u003c\/th\u003e\n\u003ctd\u003e100 assays\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eKit Plus (E. coli Topo I included), 100 assays (DRA100KE)\u003c\/th\u003e\n\u003ctd\u003e100 assays (96-well) or 200 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 (DRA100K)\u003c\/th\u003e\n\u003cth\u003eKit Plus (E. coli Topo I included), 100 assays (DRA100KE)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Buffer T1\u003c\/td\u003e\n\u003ctd\u003e480 µl\u003c\/td\u003e\n\u003ctd\u003e480 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\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eDye H19 (also sold separately as H19 Dye for DNA Relaxation and Supercoiling Assays, BHT16300181)\u003c\/td\u003e\n\u003ctd\u003e8 µl (4500 x)\u003c\/td\u003e\n\u003ctd\u003e20 µl (1500 x)\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\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 x E. coli topoisomerase I\u003c\/td\u003e\n\u003ctd\u003eNot included\u003c\/td\u003e\n\u003ctd\u003e42 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eFinal bacterial topo I reaction: 16 mM Tris-HCl pH 8, 6 mM MgCl2, 25 µg\/ml supercoiled plasmid DNA and 40 nM topoisomerase I. Free magnesium is needed for H19 detection, so high EDTA should be avoided in the reaction and detection solutions.\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eThe kit uses the fluorescence dye H19, which in the presence of magnesium gives a much weaker signal with relaxed DNA than with supercoiled DNA. As topoisomerase I relaxes the supercoiled plasmid, fluorescence at 535 nm (excitation 485 nm) decreases.\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\u003eTopo I DNA relaxation activity\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\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 detection)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSizes\u003c\/th\u003e\n\u003ctd\u003eKit: 100 assays; Kit Plus: 100 assays (96-well) or 200 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\u003eE. coli DNA Topoisomerase I Assay (PDF, see Documents): Covers catalog # DRA100KE.\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 DNA Topoisomerase I 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)","offer_id":67794878497133,"sku":"DRA100K","price":825.0,"currency_code":"USD","in_stock":true},{"title":"Kit Plus (E. coli Topo I included)","offer_id":67794883576173,"sku":"DRA100KE","price":1057.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/Topo_I_relaxation_assay.jpg?v=1792791968","url":"https:\/\/www.ebiohippo.com\/products\/dna-topoisomerase-i-assay-kit-bht16300061","provider":"BioHippo","version":"1.0","type":"link"}