CDK9+Cyclin K NanoBRET Kinase Assay

CDK9+Cyclin K NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target. 

Target

CDK9+Cyclin K

HGNC Symbol

CDK9-CCNK

Synonyms

C-2k, CDC2L4, PITALRE, TAK

Cell Line

HEK293 human epithelial cell line from ATCC

Assay Format

384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay

Assay Protocol

HEK293 cells transiently expressing NanoLuc-CDK9 Fusion Vector and Cyclin K Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRET Tracer K-10 and then treated with reference compound Dinaciclib 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.

Readout

Binding of compound to target kinase

Reference Compound IC50s
Compounds IC50 (M)
Dinaciclib 7.719e-009
Screening Location

Malvern, USA

Reference compound IC50 for CDK9+CycK