| Assay Method Information | |
| | Enzyme Inhibition Assay |
| Description: | The enzyme reaction was started by the addition of enzyme to the reaction mixture containing substrate and test compounds. The reaction was stopped by spotting an aliquot onto a silica gel plate that had been prespotted with thymine and thymidine. The plate was developed in ethyl acetate-water-formic acid (60:35:5), and the spots were visualized under UV light (254 nm) and cut out for radioactivity determination in toluene-based scintillation cocktail. Kinetic constants Km and Ki were determined from the Lineweaver-Burk and Dixon plots. Data based on results from at least four independent experiments were evaluated by the nonlinear regression method (GOSA, Bio-Log). |
| Affinity data for this assay | |
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