| Assay Method Information | |
| | In Vitro Protein Kinase Assay |
| Description: | NSC 109555 was diluted in water. All other drugs were dissolved in DMSO, in which case the final DMSO concentration in reactions was 10%, and the controls were performed under comparable conditions. Recombinant Chk2 was incubated with either histone H1 (or GST-Cdc25C) in reaction buffer, and incubated at 30 deg C for indicated times. For the drug inhibition experiments, samples were coincubated with drug during the reactions. Reactions were stopped by adding sample buffer, and samples were boiled for 5 min. Reaction products were separated by 4-20% SDS-PAGE. Chk2 protein kinase activity, measured as 32P incorporation into Chk2, histone H1 (or GST-Cdc25C), was determined using a PhosphorImager (GE Healthcare, Chalfont St. Giles, Buckinghamshire, UK). Densitometry was performed using Image-Quant. |
| Affinity data for this assay | |
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