| Field | Specification |
|---|---|
| Organism | |
| Applications | |
| Assay type | |
| Assay target | |
| Assay area | |
| Target type | |
| Detection method | |
| Detection wavelength | |
| Instrument required | |
| Plate format | |
| Assay time | |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
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
Can’t find the exact format you need? We can help you source or customize this E. coli gyrase ATPase activity assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.