Glycogen Synthase Kinase 3 beta, Recombinant, Human (GSK3b)

Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine kinase that was first characterized by its role in intermediary metabolism. The enzyme is the mammalian homolog of the zeste-white3 (shaggy) homeotic gene of Drosophila melanogaster and has been implicated in the regulation of the c-Jun/AP-1 transcription factor. In mammals, this protein serine kinase is encoded by two related genes called GSK-3a (51kD) and b (46kD). Genetic and biochemical evidence now indicates that GSK-3 plays a role in intracellular signaling pathways. For example, during insulin and growth factor signaling the GSK-3 beta isoform becomes phosphorylated on its N-terminal serine-9 residue, presumably by either the action of MAPKAP Kinase-1 (p90 S6 kinase, Rsk-2) or p70 S6 kinase. Such phosphorylation of GSK-3 blocks its ability to phosphorylate c-Jun, protein phosphatase inhibitor 2 or the GSK-3 substrate peptide.\n\nQuality Control:\nMS Tryptic Fingerprint: Confirmed identity as GSK3b with 52% amino acid coverage of the translated native sequence listed in the protocol.\nGSK3b Kinase Assay: 2-6ng phosphorylated 20uM Glycogen Synthase Peptide 2, Phosphorylated (Substrate) (G8170-26) in the assay described in protocol.\n\nHandling Recommendations:\nRapidly thaw the vial under cold water. Immediately place on ice. Aliquot unused material into pre-chilled microcentrifuge tubes. Immediately snap-freeze the vials in liquid nitrogen prior to re-storage at -70 degrees C.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -70 degrees C. Aliquots are stable for at least 6 months at -70 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

G8170-46

Size

10ug

Form

Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 150mM sodium chloride, 0.1mM EGTA,\n0.03% Brij-35, 270mM sucrose, 0.2mM PMSF, 1mM benzamidine, 0.1% 2-mercaptoethanol.

Purity

Purified by Ni/NTA agarose chromatography ((same/more than) 85%, Verified by SDS-PAGE and Coomassie Blue stain).

References

1. Sutterland, C., et al., Biochem J. 296: 15 (1993). 2. Sutterland, C., et al., FEBS Lett. 338: 37 (1994). 3. Woodget, J.R., Semin. Cancer Biol. 5: 269 (1994). 4. Cross, D.A.E., et al., Nature 378: 785-789 (1995).

Additional Info

Specific Activity: ~900u/mg.\nUnit Definition: One unit equals 1nmole phosphate incorporated into 20uM Glycogen Synthase Peptide 2, Phosphorylated (Substrate) (G8170-26) per minute at 30 degrees C with a final ATP concentration of 100uM.