NanoBRET Target Engagement Intracellular Kinase Assay, Competitive binding assay format
CAK-kinase p42; CCRK; CDCH; p42; PNQALRE
HEK293 human epithelial cell line from ATCC
384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay
HEK293 cells transiently expressing NanoLuc-CDK20 Fusion Vector and Cyclin H Vector were seeded into the wells of 384-well plates. The cells were pretreated with the NanoBRET Tracer K-10 and then treated with reference compound PBI7627 for 1 hour. The BRET signal was measured on an Envision 2104 Multilabel Reader. IC50 value was calculated and IC50 curve was plotted using the GraphPad Prism 4 program based on a sigmoidal dose response equation.
Binding of compound to target kinase