HEK293 human epithelial cell line from ATCC
384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay
HEK293 cells transiently co-expressing NanoLuc-CDK5 fusion vector and CDK5R1 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 APY-69 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