S. aureus DNA Helicase ATPase assay Kit Plus

Mfr.No.	DNAB100KS, DNAB500KS
Description	DNA helicase (DnaB) hydrolyzes ATP as the source of molecular energy to carry out DNA unwinding required by the DNA replication process. Inhibition of the ATPase activity of DNA helicase blocks its DNA unwinding function. The DNA helicase ATPase assay can be used for high-throughput screen of DNA helicase inhibitors in drug discovery. The DNA Helicase 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 S. aureus DNA Helicase ATPase assay Kit Plus-100 (Catalog No. DNAB100KS) includes 500 µl of 10 x assay buffer, 35 µl of 100 x DNA, 150 µl of 20 x S. aureus DNA helicase, 35 µl of 100 x ATP and 5 ml of dye for 100 assays in a 384-well assay format.
Drug target assays	Antibacterial drug targets

Product Manual	Datasheet
Protocols Description	Assay Protocol 1.Reagent preparation: For each 10 assay reactions, prepare (1) 297µl of premix composed of 247.5µl of H2O, 33µl of 10 x Buffer and 16.5µl of 20 x S. aureus DNA helicase; (2) 33µl of 10 x Enzyme substrate by mixing 3.3µl of 100 x ATP and 3.3µl of 100 x DNA and 26.4µl of water. 2. Reaction: Mix 27µl of the premix with 3µl of the 10 x Enzyme substrate in each well. Incubate the reaction mixture at 37°C for 90 min. 3. Detection: Add 45µl of the Dye MPA3000 into the 30µl of the reaction mixture. Incubate for 5 min. Measure the light absorbance at 650 nm. Note: The final concentrations for the ATPase assays of the helicases are 20 mM HEPES, pH 7.5, 20 mM potassium glutamate, 1 mM DTT, 0.005% Triton X-100, 10 mM MgCl2, 20µg /ml DNA, 0.25 mM ATP and 2 µM DNA helicase. A negative control reaction can be the reaction mixture without addition of ATP or enzyme.

Have you used this product?

Combination SKU	Supplier No.	Size	Your price	Quantity
BHE16300069-1	DNAB100KS	100 assays	$634.60		Add to Cart
BHE16300069-2	DNAB500KS	500 assays	$2025.58		Add to Cart