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.
Applications:Suitable for use in Western Blot.
Not suitable for use in Immunoprecipitation.
Other applications have not been tested.
Recommended Dilutions: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.
Included Positive Antigen Control:J9000: Jurkat lysate.
Use 20ug per lane for minigels.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size:200ug
Host:mouse
Source Antibody:drosophila
Grade:Affinity Purified
Purity:Purified by Protein G affinity chromatography from ascites.
Form:Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide.
Specificity:Recognizes Drosophila Glycogen Synthase Kinase 3, phosphorylated (Y279/Y216) (phosphoGSK-3) at 46kD (alpha) and 51kD (beta). Species Crossreactivity: mouse, rat and human. Expected to crossreact with nearly all species because of the extremely high conservation of this region among the known enzymes of many different species.
Clone # USB:0.T.57
Isotype:IgG1,k
Calc Applications Abbrev:WB
Calc Crossreactivity:Dr Hu Mo Rt
Immunogen:Synthetic peptide corresponding to aa203-219, C-KQLLHGEPNVS[pY]ICSRY, from the catalytic domain (between subdomain VII & VIII) of the Drosophila GSK-3/shaggy enzyme coupled to ovalbumin.