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| | CDK1 Inhibition Assay |
| Description: | CDK1 kinase activity was measured in a DELFIA based HTScan CDK1/CycB kinase assay kit (Cell Signalling Technology) that monitors biotinylated substrate peptide using a specific phospho antibody. The enzyme reaction was carried out in polypropylene plates (Greiner). The reaction mix contained CDK1/CycB kinase, Rb (Ser780) biotinylated peptide, ATP, and test compound diluted to a give a range of concentrations. The reaction was incubated for 30 min at room temperature and stopped. An aliquot of the reaction mix was transferred to a yellow streptavidin coated plate. The primary antibody Phospho Rb (Ser 780), and subsequently europium-labelled anti-rabbit IgG were added. The plate was read on an Envision 2103 multilabel counter (PerkinElmer Life Sciences) using a time-resolved measurement mode reading fluorescence at 615 nm. |
| Affinity data for this assay | |
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