OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Glycogen synthase kinase 3 (GSK3) is a protein kinase that was originally identified as a regulator of glycogen synthase, a key enzyme in glycogen metabolism. There exists two isoforms of GSK3, GSK3A and GSK3B, and they are strictly regulated via phosphorylation. GSK3 has been shown to phosphorylate a number of different cellular proteins, including p53, c-Myc, c-Jun, heat shock factor-1 (HSF-1), and cyclin D1. GSK3A also has been inhibited by activation of the canonical Wnt pathway, resulting in increased catenin that drives human mesenchymal stem cells to osteoblasts.
Family:
Kinase
Sub Family:
CMGC
Class:
Protein Ser/Thr
Protein Name:
GSK3alpha
Uniprot Number:
P49840
Protein Aliases:
Serine/threonine-protein kinase GSK3A
Gene Name:
GSK3A
Gene ID:
2931
Accession Number:
NM_019884
Organism:
Human
Construct Details:
2-end; S449A recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
GSK3a (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 20 µM YRRAAVPPSPSLSRHSSPHQS(p) EDEEE (phospho GS2 peptide), 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:
20 µM YRRAAVPPSPSLSRHSSPHQpSEDEEE (phospho GS2 peptide)
Tracer:
33P
ATP Concentration:
[ATP]= 10µM (Km= 10µM )
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:
Metabolic Diseases
Adverse / Beneficial Action:
Inhibitors may have therapeutic potential for the treatment of diabetes, Alzheimer's disease, osteoblast differentiation, stroke and bipolar disorder.
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
RELATED SOLUTIONS
GSK3alpha (GSK3A) Human CMGC Kinase Enzymatic Radiometric [Km ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-492KP
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