| Assay Method Information | |
| | In Vitro Kinase Detection |
| Description: | Kinases BTK wt (Carna, Cat. No 08-180) and BTK C481S (Carna, Cat. No 08-547) were prepared into a 2.5× kinase solution, and the substrates FAM-P2 (GL Biochem, Cat. No. 112394) and ATP ((Sigma, Cat. No. A7699-4G) were prepared into a 2.5× substrate solution, respectively. 5 μL of the compounds at different concentrations were added to a 384-well plate, and 10 μL of 2.5× kinase solution was added, the cultures were incubated at room temperature for 10 minutes. 10 μL of 2.5× substrate solution was added, and the mixture was incubated at 28° C. for an appropriate period of time. The reaction was stopped by adding 30 μL of stop solution, and the detection was carried out using Caliper EZ reader2 instrument. The IC50 value was calculated using XLFit excel add-in version 5.4.0.8 software. |
| Affinity data for this assay | |
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