{"product_id":"96-well-s-aureus-topo-iv-dna-cleavage-assay-kit-plus-bht16300080","title":"96-Well S. aureus Topo IV DNA Cleavage 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\u003eBacterial gyrase, mammalian topoisomerase II and bacterial topoisomerase IV (parC-parE) are all type II topoisomerases. They alter DNA topology by cutting both strands and joining them again.\u003c\/p\u003e\n\u003cp\u003eSome inhibitors, such as fluoroquinolones, stop the rejoining step. Cut (linear) DNA then stays bound to the enzyme as a cleavage complex, which damages DNA in cells (a topoisomerase poison). Checking for this complex helps show how a new inhibitor acts: ciprofloxacin and novobiocin both inhibit gyrase, 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\u003eT4C96KS\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\u003ctr\u003e\n\u003ctd\u003e100 x S. aureus Topo IV\u003c\/td\u003e\n\u003ctd\u003e26 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eA plate filtration device and a vacuum line are needed for the filtration step (not included).\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eLinear DNA passes through a solution matrix and a filter much more easily than closed circular DNA. After the reaction, the sample is treated with SDS and proteinase K, mixed with TDC matrix and filtered on the TDC filter plate. DNA collected in the black receiver plate is stained with dye and read by fluorescence (Ex 485 nm \/ Em 535 nm), so the signal rises with cleavage complex formation.\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\u003eS. aureus Topo IV DNA cleavage\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eTarget Type\u003c\/th\u003e\n\u003ctd\u003eDNA Topoisomerase IV\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eOrganism\u003c\/th\u003e\n\u003ctd\u003eS. aureus\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\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\u003eEquipment Needed\u003c\/th\u003e\n\u003ctd\u003eLab vacuum line or pump (up to 80 kPa or 600 mmHg), plate vacuum device, plate fluorescence reader (Ex 485 nm \/ Em 535 nm)\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 # T4C96KS.\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 S. aureus Topo IV DNA Cleavage Assay Kit Plus 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":67794879152493,"sku":"T4C96KS","price":1313.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/TDC_fig_amall_7fb7683d-1947-4876-89b2-421ed95eca4b.jpg?v=1792791969","url":"https:\/\/www.ebiohippo.com\/products\/96-well-s-aureus-topo-iv-dna-cleavage-assay-kit-plus-bht16300080","provider":"BioHippo","version":"1.0","type":"link"}