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DNA decatenation is an essential step during DNA replication in cells. In humans it is carried out by topoisomerase II, which converts concatenated DNA into decatenated DNA.
Kit Components
| Component | HDD96KE |
|---|---|
| 10 x Assay buffer | 600 µl |
| 50 mM MgCl2 | 1200 µl |
| 10 x Concatenated DNA (30 µg/ml kDNA) | 510 µl |
| 10 x ATP (10 mM) | 520 µl |
| 0.5 M EDTA | 600 µl |
| 20 x Fluorescence dye | 275 µl |
| Human topoisomerase II alpha (10 U/µl) | 50 µl |
| 10 x Rinse buffer | 2 ml |
| V-bottom plate | 1 plate |
| TDD filter plate | 1 plate |
| Black 96-well plate | 1 plate |
Assay Principle
After the topo II reaction, decatenated DNA is separated from concatenated kDNA by vacuum filtration through a TDD filter plate. The decatenated DNA passes into a black 96-well receiver plate and is quantified with a fluorescence dye at 535 nm (excitation 485 nm).
Specifications
| Assay Target | Human topo II DNA decatenation |
|---|---|
| Target Type | Topoisomerase |
| Organism | Human |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | 96-well filter plate |
| Instrument Required | Fluorescence microplate reader |
| Equipment Required | Lab vacuum line or pump (up to 80 kPa / 600 mmHg) and a plate vacuum device (not provided) |
| Size | 96 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- 96-Well Human Topo II DNA Decatenation Assay (PDF, see Documents): Covers catalog # HDD96KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this human topo II DNA decatenation 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.