| Assay Method Information | |
| | Di-UB K63-2 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. Di-Ub K63-2 hydrolysis assay was performed in 50 mM Tris/HCl pH=7.5, 1mM DTT. USP2a (20 nM) was preincubated with an excess of tested compound for 30 min. Fluorescence changes were recorded at excitation and emission wavelengths of 540 nm and 580 nm respectively, immediately after Di-Ub (200 nM, LifeSensors) 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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