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E. coli DNA Helicase ATPase assay Kit Plus

BHE16300063

This product includes 0.500 ml of 10 x assay buffer, 0.035 ml of 100 x DNA, 0.030 ml of 100 x E. coli DNA helicase, 0.035 ml of 100 x ATP and 5 ml of dye for 100 assays in a 384-well assay format.

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+1 866.986.9598
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[email protected]
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Reference Nakano T., et al, Translocation and Stability of Replicative DNA Helicases upon Encountering DNA-Protein Cross-links, J. Biol. Chem. 288: 4649-4658 (2013). 
Product Manual Datasheet
Protocols Description Assay Protocol
1.Reagent preparation:
For each 10 assay reactions,
(1) Prepare 297µl of premix composed of 261µl of H2O, 33µl of 10 x Buffer and 3.3µl of 100 x E. coli DNA helicase.
(2) Prepare 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 60 min.
 
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, 250 nM DNA, 0.25 mM ATP and 200 nM DNA helicase. A negative control reaction can be the reaction mixture without addition of ATP or enzyme.
 
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.
Combination SKU Supplier No. Size Your price Quantity
BHE16300063-1 DNAB100KE 100 assays $613.14
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BHE16300063-2 DNAB500KE 500 assays $1957.08
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