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. 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. GSK-3 is constitutively phosphorylated at this tyrosine residue in resting cells. This immunogen is identical in 15 of 17 residues in human and rat GSK-3a (Y279) and GSK-3b (Y216). The phosphotyrosine residue can be removed from GSK-3 by treatment with PTP1B.

Suitable for use in Western Blot. Not suitable for use in Immunoprecipitation. Other applications have not been tested.

Western Blot: 0.5-2ug/ml detects phospho-GSK-3 in RIPA lysates from human Jurkat cells. Previous lots detected phospho-GSK-3 in mouse 3T3/A31, human A431 and rat L6 cell lysates. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with G8170-47. Proteins were visualized using an IgG (HRP) goat anti-mouse and a chemiluminescence detection system Optimal dilutions to be determined by the researcher.

J9000: Jurkat lysate. Use 20ug per lane for minigels.

Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight™650 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Note: Applications are based on unconjugated antibody.

Synthetic peptide (aa203-219, C-KQLLHGEPNVS[pY]ICSRY) corresponding to the catalytic domain (between subdomain VII & VIII) of Drosophila GSK-3/shaggy enzyme coupled to ovalbumin (Accession# NM_057366).

Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™650.

Purified by Protein G affinity chromatography.

Recognizes Drosophila Glycogen Synthase Kinase 3, phosphorylated (Y279/Y216) (phosphoGSK-3) at 46kD (alpha) and 51kD (beta). Species Crossreactivity: mouse, rat, human, S. pombe and Dictyostelium. Expected to crossreact with nearly all species because of the extremely high conservation of this region among the known enzymes of many different species.