| Assay Method Information | |
| | In-Vitro Kinase Activity Assay |
| Description: | Kinase reactions are assembled in 384-well plates (Greiner) in a total volume of 20 μL. Test compounds (3a-3g) were diluted in DMSO to a final concentration, while the final concentration of DMSO in all assays was kept at 1%. The compounds were incubated with the kinases for 30 min. A 0.5 nM concentration of BTK in 100 mM HEPES, pH 7.5; 0.1% BSA, 0.01% Triton X-100, 1 mM DTT, 5 mM MgCl2, were used. The reaction was initiated by 2-fold dilution into a solution containing 5 μM ATP and 1 μM substrate in the kinase buffer. |
| Affinity data for this assay | |
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