		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. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Type	Isotype	Clone	Grade
Mab	IgG1	0.T.56	Affinity Purified

Size		Storage	Shipping	Source	Host
100ug		-20°C	Blue Ice	Drosophila	Mouse

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 (ovalbumin).

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.