Technical Data
G8170-40
Glycogen Synthase Kinase 3 (GSK3)
Description:
Glycogen synthase kinase-3 (GSK-3), a multifunctional serine/threonine kinase, is a key regulator of numerous signaling pathways. Two isoforms of GSK-3 are reported in mammals: a 51kD GSK-3a and a 47kD GSK-3b. These two isoforms exhibit about 98% homology in their kinase domains, but share only about 36% identity in the last 76 C-terminal amino acid residues. GSK-3a contains a glycine-rich extension at its N-terminus. A minor (~15% of total) splice variant of GSK-3b, GSK-3b2, has also been identified, which contains a 13-residue insert within the kinase domain. It exhibits reduced kinase activity towards tau protein compared with ‘unspliced’ GSK-3b. GSK-3b2 is localized primarily to neuronal cell bodies, unlike unspliced GSK-3b that is also found in neuronal processes. GSK-3b plays a key inhibitory role in the Wnt signaling pathway. Wnt genes encode a large family of secreted, cysteine-rich proteins that are important in development and in maintenance of adult tissues. Abnormalities in Wnt signaling are reported to promote both human dgenerative diseases and cancer.

Applications:
Suitable for use in Western Blot and Immunocytochemistry. Not suitable for use in Immunoprecipitiation. Other applications not tested.

Recommended Dilution:
Western Blot: 0.5-2ug/ml. Detects GSK-3 in RIPA lysates from human Jurkat and A431, rat L6 and PC12 and mouse 3T3 cells. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with G8170-40 (0.5ug/ml). Proteins were visualized using an IgG (HRP) goat anti-mouse and a chemiluminescence detection system.
Immunocytochemistry: 10ug/ml. Demonstrated positive immunostaining for GSK3 in L6 cells fixed with acetone.
Optimal dilutions to be determined by the researcher.

Positive Control (Included): J9000: Jurkat Cell

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.    
TypeIsotypeCloneGrade
MabIgG10.T.56Affinity Purified
SizeStorageShippingSourceHost
100ug-20°CBlue IceDrosophilaMouse
Concentration:
~1mg/ml
Immunogen:
Synthetic peptide corresponding to aa203-219 (18aa, CKQLLHGEPNVSYICSRY) derived from a region of the catalytic domain (between subdomain VII and VIII) of the Drosophila GSK-3/ shaggy enzyme, coupled with ovalbumin (NP_476714)
Purity:
Purified by Protein G affinity chromatography.
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 alpha (51kD) and beta (46kD) isoforms. Due to extremely high conservation among the known enzymes of many different species, should crossreact with nearly all species homologues. Species Crossreactivity: mouse, rat, S. pombe, Dictyostelium and human.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
Hughes, K., et al., Eur. J. Biochem. 203: 305-311, 1992. Woodgett, J.R., et al., EMBO 12: 803-808, 1993. Siddle, K., et al., Biochem J. 305: 25-28, 1995. Application Harwood, et al.; Cell 80: 139-148, 1995.