E. coli DNA Helicase ATPase Assay Kit Plus

SKU:BHT16300046
Overview
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E. coli DNA Helicase ATPase Assay Kit Plus, a colorimetric kit for the ATPase activity of E. coli DnaB helicase and for helicase inhibitor screening. Phosphate released is read at 650 nm in a 384-well plate. Comes with enzyme, DNA and ATP; 100 or 500 assays.
Assay Target E. coli DNA helicase ATPase
Detection Method Colorimetric
Detection Wavelength Absorbance 650 nm
Plate Format 384-well plate
Assay Time About 65 min
Options selector
Catalog no. Size
DNAB100KE 100 assays
DNAB500KE 500 assays
Available Options

Select the variant that best fits your experiment. Availability and lead time may vary by option.

  • Options: Size: 100 assays, 500 assays
  • Lead time: options listed in "Availability Content"; other statuses may take longer.
  • Storage: Please contact us to confirm storage conditions.
  • Shipping: Shipped frozen.
  • Upon receipt: store at recommended temperature as soon as possible.
  • Sales terms and conditions: Please review prior to ordering.
Field Specification
Organism E. coli
Applications
  • Enzyme Activity
  • Compound Profiling
Assay type
  • Enzyme Activity Assay
  • Inhibitor Screening Assay
Assay target E. coli DNA helicase ATPase
Assay area Antibacterial Drug Discovery
Target type Helicase
Detection method
  • Colorimetric
Detection wavelength Absorbance 650 nm
Instrument required Absorbance microplate reader
Plate format 384-well plate
Assay time
  • About 65 min
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.
Catalog no. (Mfr.) DNAB100KE | DNAB500KE
Main SKU BHT16300046
Drug Target Assay Kit

During DNA replication, DNA helicase (DnaB) unwinds DNA using energy from ATP hydrolysis. When the helicase ATPase is blocked, DNA unwinding stops too, so this ATPase reaction serves as a drug discovery screen for helicase inhibitors.

Pack Sizes

Both sizes include the E. coli DNA helicase. The only difference is the number of assays: DNAB100KE provides reagents for 100 assays and DNAB500KE for 500 assays (384-well format), with component volumes scaled accordingly.

100 assays (DNAB100KE) 100 assays; enzyme included
500 assays (DNAB500KE) 500 assays; enzyme included

Kit Components

Component 100 assays (DNAB100KE) 500 assays (DNAB500KE)
10 x Assay buffer 500 µl 2 ml
100 x DNA 35 µl 170 µl
100 x E. coli DNA helicase 30 µl 150 µl
100 x ATP 35 µl 170 µl
Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) 5 ml 25 ml

Final reaction conditions: 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.

Assay Principle

The kit detects the phosphate produced when E. coli DNA helicase hydrolyzes ATP in the presence of DNA. A phosphate detection dye is added and absorbance is read at 650 nm; inhibitors of the helicase ATPase lower the signal.

Specifications

Assay Target E. coli DNA helicase ATPase
Target Type Helicase
Organism E. coli
Assay Type Enzyme Activity Assay; Inhibitor Screening Assay
Detection Method Colorimetric
Detection Wavelength Absorbance 650 nm
Plate Format 384-well plate
Instrument Required Absorbance microplate reader
Assay Time About 65 min (60 min reaction + 5 min detection)
Publication Nakano T. et al., J. Biol. Chem. 288: 4649-4658 (2013)
Sizes 100 assays, 500 assays
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.

Protocol

The manufacturer's protocol describes the procedure and reaction volumes.

  • E. coli DNA Helicase ATPase assay (PDF, see Documents): Covers catalog # DNAB100KE, DNAB500KE.

Safety & Handling

For Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The E. coli DNA Helicase ATPase Assay Kit Plus is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Please contact us to confirm storage conditions.
Q.What does this kit measure?
A.It measures the DNA-dependent ATPase activity of E. coli DNA helicase by detecting the phosphate released from ATP hydrolysis. It is used for high-throughput screening of DNA helicase inhibitors.
Q.What is the difference between the options?
A.Both sizes include the E. coli DNA helicase. The only difference is the number of assays: DNAB100KE provides reagents for 100 assays and DNAB500KE for 500 assays (384-well format), with component volumes scaled accordingly.
Q.Is a protocol available?
A.Yes. The manufacturer's protocol (E. coli DNA Helicase ATPase assay, linked under Documents) covers catalog # DNAB100KE, DNAB500KE.
Q.How long does the assay take?
A.About 65 min (60 min reaction + 5 min detection). See the protocol for the exact steps.
Q.How should the kit be stored?
A.The E. coli DNA Helicase ATPase Assay Kit Plus is shipped frozen. Please contact us for long-term storage conditions.

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