{"product_id":"e-coli-topo-iv-atpase-assay-kit-plus-bht16300084","title":"E. coli Topo IV ATPase Assay Kit Plus","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\u003eType II DNA topoisomerases use ATP hydrolysis to drive DNA supercoiling (gyrase) and decatenation (topoisomerase IV). Blocking this ATPase activity, as novobiocin does, also blocks the enzyme's biological role, which makes the E. coli Topo IV ATPase reaction a useful readout for screening topoisomerase inhibitors.\u003c\/p\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\u003eT4A-100KE\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Buffer\u003c\/td\u003e\n\u003ctd\u003e400 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 x DNA\u003c\/td\u003e\n\u003ctd\u003e35 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 x E. coli Topo IV\u003c\/td\u003e\n\u003ctd\u003e33 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 x ATP\u003c\/td\u003e\n\u003ctd\u003e35 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003ePhosphate detection dye\u003c\/td\u003e\n\u003ctd\u003e5 ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eThe 100 x DNA stock is 1 mg\/ml. Final reaction conditions: 20 mM Tris-HCl pH 8, 35 mM NH4OAc, 4.6% glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 10 µg\/ml DNA, 0.2 mM ATP and 50 nM topoisomerase.\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eThe kit detects the inorganic phosphate released when topoisomerase IV hydrolyzes ATP in the presence of DNA. A phosphate detection dye is added after the reaction and absorbance is read at 650 nm; inhibitors lower the signal.\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\u003eE. coli Topo IV ATPase 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\u003eOrganism\u003c\/th\u003e\n\u003ctd\u003eE. coli\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Type\u003c\/th\u003e\n\u003ctd\u003eEnzyme Activity Assay; Inhibitor Screening Assay\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eDetection Method\u003c\/th\u003e\n\u003ctd\u003eColorimetric\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eDetection Wavelength\u003c\/th\u003e\n\u003ctd\u003eAbsorbance 650 nm\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003ePlate Format\u003c\/th\u003e\n\u003ctd\u003e384-well plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eInstrument Required\u003c\/th\u003e\n\u003ctd\u003eAbsorbance microplate reader\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Time\u003c\/th\u003e\n\u003ctd\u003eAbout 60 min\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSize\u003c\/th\u003e\n\u003ctd\u003e100 assays\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\u003eDNA Topoisomerase ATPase Assays (PDF, see Documents): Covers catalog # T4A-100KE.\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 E. coli Topo IV ATPase Assay Kit Plus 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":"100 assays","offer_id":67794878857581,"sku":"T4A-100KE","price":829.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/topo_ATPase_assay_64e24412-be6d-48b6-8dbc-43a905dfd3c6.jpg?v=1792791969","url":"https:\/\/www.ebiohippo.com\/products\/e-coli-topo-iv-atpase-assay-kit-plus-bht16300084","provider":"BioHippo","version":"1.0","type":"link"}