| Assay Method Information | |
| | Inhibition Assay |
| Description: | This example demonstrates reversible inhibition of USP1/UAF1 by an embodiment of the invention.The compound (81) at a concentration of ten times its 1050 value was pre-incubated with 10 μM USP1/UAF1 at room temperature for 15 min. This solution was then diluted by 100-fold in assay buffer containing 50 mM HEPES, pH 7.8, 0.1 mg/mL BSA, 0.5 mM EDTA, and 1 mM DTT and incubated at room temperature for an additional 15 min. Next, 20 μM K63-linked diubiquitin was added into the solution and incubated at 37° C. for 10 min. Laemmli sample buffer was added to quench the reaction. The reaction products were then separated on a 20% denaturing SDS-PAGE gel and stained with Coomassie Blue. USP1/UAF1 incubated with DMSO was treated as 100% USP1/UAF1 activity. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |