CDK4+Cyclin D1 NanoBRET Kinase Assay

CDK4+Cyclin D1 NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.
Target Overview


CDK4+Cyclin D1

HGNC Symbol

CDK4/CCND1 complex


CMM3; PSK-J3; MGC14458

Cell Line

HEK293 human epithelial cell line from ATCC

Assay Properties

Assay Format

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

Assay Protocol

HEK293 cells transiently co-expressing NanoLuc-CDK4 fusion vector and Cyclin D1 Vector were seeded into the wells of 384-well plates. The cells were pretreated with the NanoBRET Tracer K-7 and then treated with reference compound Palbociclib 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

Reference Compound IC50s
Compounds IC50 (M)
Palbociclib 4.028e-009

Screening Location

Malvern, USA

Reference compound IC50 for CDK4+Cyclin D1