{"product_id":"96-well-topoisomerase-dna-cleavage-assay-kit-bht16300111","title":"96-Well Topoisomerase DNA Cleavage 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\u003eType II topoisomerases, which include mammalian topoisomerase II, bacterial gyrase and topoisomerase IV (parC-parE), alter DNA topology by cutting both strands and joining them again. If a compound such as a fluoroquinolone stops the rejoining, the cut (linear) DNA stays covalently linked to the enzyme as a cleavage complex, which harms DNA in cells (a topoisomerase poison).\u003c\/p\u003e\n\u003cp\u003eChecking for cleavage complex formation helps show how a new topoisomerase inhibitor acts. Novobiocin and ciprofloxacin both inhibit DNA gyrase, for example, yet only ciprofloxacin leads to cleavage complexes.\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\u003eTDC96K\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Reaction buffer\u003c\/td\u003e\n\u003ctd\u003e300 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x DNA\u003c\/td\u003e\n\u003ctd\u003e30 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eSDS solution\u003c\/td\u003e\n\u003ctd\u003e260 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 mM ATP\u003c\/td\u003e\n\u003ctd\u003e30 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eProteinase K solution\u003c\/td\u003e\n\u003ctd\u003e260 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eTDC matrix\u003c\/td\u003e\n\u003ctd\u003e10 ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eRinse buffer\u003c\/td\u003e\n\u003ctd\u003e6 ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Dye\u003c\/td\u003e\n\u003ctd\u003e500 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eBlack receiver plate\u003c\/td\u003e\n\u003ctd\u003e1 plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eV-bottom reaction plate\u003c\/td\u003e\n\u003ctd\u003e1 plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eTDC filter plate\u003c\/td\u003e\n\u003ctd\u003e1 plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eTopoisomerase is not included. A plate vacuum filtration device and a lab vacuum line (up to 80 kPa or 600 mmHg) are needed and are not provided.\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eAfter the topoisomerase reaction, the protein is removed with SDS and proteinase K and the sample is mixed with a matrix. Linearized DNA passes through the matrix and filter plate much more easily than closed circular DNA; the filtered DNA is collected in a black plate, stained with a dye and read at 535 nm (excitation 485 nm), so cleavage-complex formers raise 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\u003eTopoisomerase DNA cleavage complex\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\u003eInhibitor Screening 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 filter plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eInstrument Required\u003c\/th\u003e\n\u003ctd\u003eFluorescence reader + vacuum device\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Time\u003c\/th\u003e\n\u003ctd\u003eAbout 130 min (60 min reaction + 60 min proteinase K + 5 min matrix mix + about 5 min filtration)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eEquipment Needed\u003c\/th\u003e\n\u003ctd\u003eLab vacuum line or pump (up to 80 kPa \/ 600 mmHg), plate vacuum device, fluorescence plate reader\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSize\u003c\/th\u003e\n\u003ctd\u003e96 assays\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eStorage\u003c\/th\u003e\n\u003ctd\u003eStore the TDC filter plate and TDC matrix at 4 °C and the other reagents at -20 °C.\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\u003eTopoisomerase DNA Cleavage Assays (PDF, see Documents): Covers catalog # TDC96K.\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 96-Well Topoisomerase DNA Cleavage Assay Kit is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Store the TDC filter plate and TDC matrix at 4 °C and the other reagents at -20 °C.\u003c\/div\u003e\n\u003c\/div\u003e","brand":"ProFoldin","offers":[{"title":"96 assays","offer_id":67794880201069,"sku":"TDC96K","price":905.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/TDC_fig_amall_00f21ad0-30a7-4d05-9534-0233110a72b1.jpg?v=1792791969","url":"https:\/\/www.ebiohippo.com\/products\/96-well-topoisomerase-dna-cleavage-assay-kit-bht16300111","provider":"BioHippo","version":"1.0","type":"link"}