| Assay Method Information | |
| | CHK Kinase Inhibition Assay |
| Description: | CHK kinase function was measured in a DELFIA assay in order to monitor phosphorylation of a CDC25C peptide using a specific phospho antibody. The enzyme reaction was carried out in polypropylene plates (Greiner) using a reaction mixture containing enzyme, substrate peptide and ATP in the presence of test compound. After incubation, an aliquot of the stopped reaction mixture was transferred to a black neutravidin-coated plate (Perbio) and incubated for 1 h at room temperature. The plates were washed, and an antibody mixture consisting of anti-phospho CDC25C and europium-labelled anti-rabbit IgG was added. The plate was read on a Victor2 1420 multilabel counter (Perkin Elmer Life Sciences) using a time-resolved measurement mode reading fluorescence at 615 nm. The concentration of test compound required to inhibit enzyme activity by 50% was calculated (IC50). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |