| Assay Method Information | |
| | Assay Condition A |
| Description: | The IC50 profile of test compounds was determined using three protein kinases. IC50 values were measured by testing 10 concentrations (1×10−04M, 3×10−05M, 1×10−05M, 3×10−06M, 1×10−06M, 3×10−07M, 1×10−07M, 3×10−08M, 1×10−08M, and 3×10−09M) of each compound in singlicate.Test compounds: The compounds were provided as pre-weighed powders in vials. The compounds were dissolved to 1×10−02M by adding DMSO. 100 μl of each of the resulting stock solutions were transferred into column 2 of four 96 well master plates.Prior to testing, the 1×10−02M stock solutions in column 2 of the master plates were subjected to a serial, semi-logarithmic dilution using 100% DMSO as a solvent. This resulted in 10 distinct concentrations, with a dilution endpoint of 3×10−07M/100% DMSO in column 12. Column 1 and 7 were filled with 100% DMSO as controls. Subsequently, 2×10 μl from each well of the serial diluted copy plates were aliquoted with a 96 channel pipettor into two identical sets of compound dilution plates.In the process, 90 μl H2O were added to each well of a set of compound dilution plates. To minimize potential precipitation, the H2O was added to each plate only a few minutes before the transfer of the compound solutions into the assay plates. Each plate was shaken thoroughly, resulting in a compound dilution plate/10% DMSO.For the assays, 5 μl solution from each well of the compound dilution plates/10% DMSO were transferred into the assay plates. The final volume of the assay was 50 μl. All compounds were tested at 10 final assay concentrations in the range from 1×10−04M to 3×10−09M, in singlicate. The final DMSO concentration in the reaction cocktails was 1% in all cases. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |