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DNA relaxation is the reaction in which human DNA topoisomerase I (topo I) turns supercoiled circular DNA into its relaxed form.
Kit Components
| Component | GHRA100KE |
|---|---|
| 10 x Buffer HT1 | 400 µl |
| Supercoiled DNA (0.25 mg/ml) | 100 µl |
| 5 x Gel loading buffer | 550 µl |
| Human topoisomerase I (500 U/µl) | 10 µl |
Final reaction concentrations: 25 mM Tris-Cl, pH 7.4, 57 mM KCl, 5 mM MgCl2, 0.25 mM DTT, 0.25 mM EDTA, 15 µg/ml BSA, 12.5 µg/ml supercoiled plasmid DNA and 2.5 U/µl human topo I.
Assay Principle
Supercoiled and relaxed DNA run differently on an agarose gel. After the topo I reaction, samples are separated by agarose gel electrophoresis, stained with ethidium bromide and photographed under UV light; more relaxed DNA means higher topo I activity.
Specifications
| Assay Target | Human topo I DNA relaxation |
|---|---|
| Target Type | Topoisomerase |
| Organism | Human |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Gel Electrophoresis |
| Instrument Required | UV gel imaging system |
| Equipment Required | Agarose gel electrophoresis setup and UV light for imaging |
| 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.
- Gel-Based Human Topoisomerase I DNA relaxation Assay (PDF, see Documents): Covers catalog # GHRA100KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this human topo I DNA relaxation 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.