| Assay Method Information | |
| | Ub-AMC-Hydrolysis Assay |
| Description: | The assays were performed using Infinite 200 PRO - Tecan plate reader and 96-well, black Greiner microplates in a 100 ml reaction volume. The assays were performed using Infinite 200 PRO - Tecan plate reader and 96-well, black Greiner microplates in a 100 ml reaction volume. Ub-AMC-hydrolysis assay was performed in a reaction buffer (50 mM Tris/HCl, pH=7.5, 1mM EDTA, 1mM MgCl2). USP2a (10 nM) was preincubated with an excess of tested compound for 30 minutes and fluorescence changes were recorded at excitation and emission wavelengths of 355 nm and 460 nm respectively, immediately after Ub-AMC (500 nM, Viva Biosience) addition. The excess of the wild type ubiquitin or NSC 632839 was used as the inhibition control. |
| Affinity data for this assay | |
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