DNA Topoisomerase ATPase Assay Kits

Mfr.No.	T2A-100KE, T2A-100KS, T4A-100KE, T4A-100KS, T4A-100KN
Description	Type II DNA topoisomerases including topoisomerase II and topoisomerase IV hydrolyze ATP as the source of molecular energy to carry out DNA supercoiling and DNA decatenation reactions. Inhibition of the ATPase activity of topoisomerases by compounds such as novobiocin blocks their biological functions. Therefore, the topoisomerase ATPase assay can be used for high-throughput screen of topoisomerase inhibitors. The Topoisomerase ATPase Assay Kit is based on detection of the phosphate produced by the ATP hydrolysis reaction in the presence of DNA. The assay is in a 384-well plate format and the phosphate is detected using light absorbance at 650 nm. The E. coli gyrase ATPase assay Kit Plus (Catalog No. T2A-100KE) includes 400 µl of 10 x Buffer, 35 µl of 100 x DNA, 33 µl of 100x E. coli gyrase, 35 µl of 100 x ATP and 5 ml of Dye for 100 assays of E. coli gyrase ATPase activity in a 384-well assay format.
Drug target assays	Antibacterial drug targets

Product Manual	Datasheet
Protocols Description	Assay Protocol 1.Reaction and sample preparation: The total volume of each reaction mixture is 30µl including: 18µl of H2O, 3µl of 10 x buffer, 3µl of 10 x DNA, 3µl of 10 x enzyme, 3µl of 2 mM ATP. Incubate the reaction mixture at 37°C for 60 min. Note: The 10 x DNA is prepared by 10-fold dilution of the 100 x DNA stock (1 mg/ml) in water. The 10 x enzyme is prepared by dilution of the 100 x enzyme stock in the 1x buffer. The 2 mM ATP is prepared by 50-fold dilution of the 100 mM ATP stock in water. The final concentrations are 20 mM Tris-HCl, pH 8, 35 mM NH4OAc, 4.6 % glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 10µg /ml DNA, 0.2 mM ATP and 50 nM topoisomerase. A negative control reaction can be the reaction mixture without addition of enzyme. 2. Assay Add 45ml of Dye and read the light absorbance at 650 nm.

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Combination SKU	Supplier No.	Specifications	Your price
BHE16300099-1	T2A-100KE	E. coli gyrase	$634.60	Add to Cart
BHE16300099-2	T2A-100KS	S. aureus gyrase	$634.60	Add to Cart
BHE16300099-3	T4A-100KE	E. coli Topo IV	$634.60	Add to Cart
BHE16300099-4	T4A-100KS	S. aureus Topoisomerase IV	$634.60	Add to Cart
BHE16300099-5	T4A-100KN	S. pneumoniae Topo IV	$634.60	Add to Cart