CDK4/Cyclin D1 Human CMGC Kinase Cellular TE NanoBRET LeadHunter Assay - FR
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
The NanoBRET™ Target Engagement (TE) Intracellular Kinase Assays are based on the Bioluminescence Resonance Energy Transfer (BRET) system, an energy transfer technique designed to measure molecular proximity in living cells. The assay measures the apparent affinity of test compounds by competitive displacement of the NanoBRET™ tracer, reversibly bound to a NanoLuc® luciferase-kinase fusion expressed in cells.
Family:
Kinase
Sub Family:
CMGC
Class:
Protein Ser/Thr
Protein Name:
CDK4
Uniprot Number:
P11802
Protein Aliases:
Cell division protein kinase 4|PSK-J3
Gene Name:
CDK4
Gene ID:
1019
Gene Aliases:
CMM3| PSK-J3
Accession Number:
NM_000075
Organism:
Human
Construct Details:
NanoLuc® luciferase fused to the N-terminus of the Wild Type Full-length kinase
Cell Type:
293H
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based Binding
Functional Mode:
Antagonist
Detection Method:
NanoBRET
Testing Information
Procedure Summary:
NanoBRET TE Intracellular Kinase Assays are performed according to Promega Adherent Format Assay Protocol using NanoBRET™ Tracer K-10. 293H cells are transfected with NanoLuc®-CDK4 Fusion Vector and co-transfected with CCND1 Expression Vector, seeded in 384-well plates, and cultivated overnight. Target engagement of compounds are measured by treating the cells with the compound in presence of a fixed concentration of fluorescent tracer.
Tracer:
K-10 (0.217 µM)
Incubation:
120 min at 37°C
Test Sample Requirements:
Minimum 1) Dose Response: 120 µL of 10 mM stock -OR- 1 mg (pre-weighed) for a top concentration at 30 µM)
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Additional Information
Brand:
LeadHunter
Testing Location:
France - Celle L'Evescault
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