{"product_id":"quinolone-resistant-e-coli-gyrase-mutant-s83w-assay-kit-plus-bht16300073","title":"Quinolone-resistant E. coli Gyrase Mutant S83W 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\u003eQuinolones such as ciprofloxacin are widely used antibiotics that target bacterial DNA topoisomerases. The S83W and D87G mutations in the gyrA subunit of E. coli gyrase cause quinolone resistance, so new molecules that inhibit the mutant enzymes are needed.\u003c\/p\u003e\n\u003cp\u003eThis kit measures the DNA supercoiling activity of the quinolone-resistant S83W gyrase mutant, so it can be used to test the inhibitory activity of new quinolone compounds against it.\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\u003eS83W100KE\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Buffer T2\u003c\/td\u003e\n\u003ctd\u003e600 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Relaxed DNA\u003c\/td\u003e\n\u003ctd\u003e405 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e1500 x H19 dye\u003c\/td\u003e\n\u003ctd\u003e20 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x ATP\u003c\/td\u003e\n\u003ctd\u003e450 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\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 x E. coli gyrase mutant S83W\u003c\/td\u003e\n\u003ctd\u003e50 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eFinal reaction conditions: 20 mM Tris-HCl pH 8, 35 mM NH4OAc, 4.6% glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 25 µg\/ml relaxed plasmid DNA, 1 mM ATP and 50 nM gyrase. Magnesium is essential; EDTA should be avoided.\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eRelaxed and supercoiled DNA give different fluorescence with the H19 dye: in the presence of magnesium, relaxed DNA suppresses the dye signal much more than supercoiled DNA. As gyrase converts relaxed plasmid into its supercoiled form, fluorescence (Ex 485 nm \/ Em 535 nm) rises; inhibitors keep the signal low.\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 gyrase S83W supercoiling\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eTarget Type\u003c\/th\u003e\n\u003ctd\u003eDNA Topoisomerase II (Gyrase)\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\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 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\u003eMutation\u003c\/th\u003e\n\u003ctd\u003eS83W in the gyrA subunit (quinolone-resistant)\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\u003eQuinolone-resistant E. coli Gyrase Mutant Assays (PDF, see Documents): Covers catalog # S83W100KE.\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 Quinolone-resistant E. coli Gyrase Mutant S83W 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":67794878660973,"sku":"S83W100KE","price":1283.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/mutant_gyrase_IC50_copy.jpg?v=1792791969","url":"https:\/\/www.ebiohippo.com\/products\/quinolone-resistant-e-coli-gyrase-mutant-s83w-assay-kit-plus-bht16300073","provider":"BioHippo","version":"1.0","type":"link"}