Glycogen Synthase Kinase 3 alpha, phosphorylated (Ser21) (GSK3a)

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 (51kDa) and b (46kDa). 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\nApplications: Suitable for use in Western Blotting. Immunoprecipitation (not tested). \n\nRecommended Dilution:\nWestern Blotting: 1:500-1:1000. RIPA lysates from HEK293 cells stimulated with 200ng/ml of IGF1 for 5 minutes. Non-stimulated and IGF1-stimulated HEK293 cell lysates were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-phospho-GSK3a (1:1000). Proteins, 51kD, were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 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

G8170-42B

Size

100ul

Applications

WB

Hosts

Rabbit

Reactivities

Hum, Rat

Form

Supplied as a liquid in 0.1M Tris, pH 7.4, 0.05% sodium azide, 5mg/ml BSA before the addition of glycerol to 30%.

P Type

Pab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG

References

Cross, D.A.E., et al., Nature 378: 785 789, 1995. Sutherland, C. and P. Cohen, FEBS Letter 338: 37

Additional Info

Recognizes phosphorylated human GSK-3a, Mr ~51kD, from cell lysates and phosphorylated recombinant GSK-3b. Does not recognize active dephosphorylated GSK-3a or GSK-3b. Predicted to crossreact with rat based on sequence homology (the immunizing sequence is identical in rat).