OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
CDK4 (Cyclin-dependent kinase 4) forms a complex with type D cyclins such as D1, D2 and D3 and when the cellular levels of cyclin D peak there is a shortening of the G1 phase and rapid entry into S phase of cell cycle progression. The substrate for cdk4/cyclin D is Rb, and phosphorylation of Rb results in dissociation of the E2F complex from Rb. The free E2F proteins in turn stimulate transcription of genes required for DNA synthesis. Cellular inhibition of CDK4/cyclin D3 activity is achieved by its inhibitory proteins, i.e. p16, p21 and p27. Inhibition of CDK4/CCND3 may be used in treatment of uncontrolled cell proliferation, tumor heterogeneity, invasion and metastasis.
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:
PSK-J3
Accession Number:
NM_000075.3 (CDK4)/M
Organism:
Human
Construct Details:
Full length recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
CDK4/Cyclin D3(h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.03% BSA, 0.03% Tween 20, 20 mM DTT, and 0.2 mg/ml Rb fragment, 10 mM MgAcetate and [gamma-33P-ATP] (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.
Substrate:
0.2 mg/ml Rb fragment
Tracer:
33P
ATP Concentration:
[ATP]= 10µM (Km= 408µM )
Incubation:
40 min at Room temperature
Control Inhibitor:
Staurosporine
Test Sample Requirements:
Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.
Minimum Order Quantity:
2
Turnaround Time
Standard:
6 Business Days
Service Scheduling Note:
Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
RELATED SOLUTIONS
CDK4/CyclinD3 Human CMGC Kinase Enzymatic Radiometric [Km ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-957KP
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