E. coli Gyrase ATPase Assay Kit Plus

SKU:BHT16300068
Overview
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E. coli gyrase ATPase assay kit for measuring the ATPase activity of DNA gyrase and screening inhibitors. Phosphate from ATP hydrolysis is detected with a dye at 650 nm in a 384-well format. Includes E. coli gyrase; one size of 100 assays.
Assay Target E. coli gyrase ATPase activity
Detection Method Colorimetric
Detection Wavelength Absorbance 650 nm
Plate Format 384-well plate
Assay Time About 60 min
Options selector
Catalog no. Size
T2A-100KE 100 assays
Available Options

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

  • Options: Size: 100 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 gyrase ATPase activity
Assay area Antibacterial Drug Discovery
Target type DNA Topoisomerase II (Gyrase)
Detection method
  • Colorimetric
Detection wavelength Absorbance 650 nm
Instrument required Absorbance microplate reader
Plate format 384-well plate
Assay time
  • About 60 min
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.
Catalog no. (Mfr.) T2A-100KE
Main SKU BHT16300068
Drug Target Assay Kit

Gyrase (topoisomerase II) and topoisomerase IV are type II DNA topoisomerases. They split ATP to power DNA supercoiling and decatenation. Compounds such as novobiocin that block this ATPase activity stop the enzyme from working, so an ATPase assay suits high-throughput screens for topoisomerase inhibitors.

Kit Components

Component T2A-100KE
10 x Buffer 400 µl
100 x DNA 35 µl
100 x E. coli gyrase 33 µl
100 x ATP 35 µl
Phosphate detection dye 5 ml

Final reaction conditions: 20 mM Tris-HCl pH 8, 35 mM NH4OAc, 4.6% glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 10 µg/ml DNA, 0.2 mM ATP and 50 nM topoisomerase. A reaction without enzyme can serve as negative control.

Assay Principle

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

Specifications

Assay Target E. coli gyrase ATPase activity
Target Type DNA Topoisomerase II (Gyrase)
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 60 min
Size 100 assays
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.

Protocol

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

  • DNA Topoisomerase ATPase Assays (PDF, see Documents): Covers catalog # T2A-100KE.

Safety & Handling

For Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The E. coli Gyrase 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 ATPase activity of E. coli gyrase by detecting the phosphate released when the enzyme hydrolyzes ATP in the presence of DNA. It is used to screen inhibitors that block the gyrase ATPase.
Q.What is included in the kit?
A.The E. coli Gyrase ATPase Assay Kit Plus (100 assays) includes 10 x buffer (400 µl), 100 x DNA (35 µl), 100 x E. coli gyrase (33 µl), 100 x ATP (35 µl) and phosphate detection dye (5 ml).
Q.Is a protocol available?
A.Yes. The manufacturer's protocol (DNA Topoisomerase ATPase Assays, linked under Documents) covers catalog # T2A-100KE.
Q.How long does the assay take?
A.About 60 min. See the protocol for the exact steps.
Q.How should the kit be stored?
A.The E. coli Gyrase ATPase Assay Kit Plus is shipped frozen. Please contact us for long-term storage conditions.

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Experience the power of Celltrypse™, c-LEcta's innovative enzyme solution for gentle and efficient cell dissociation. Request your free sample and discover a superior alternative for your cell culture workflows.

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