Analysis Note

ControlJurkate cell lysate, 293T cell extract, rabbit spleen fibroblasts

Included Positive Antigen Control:
Catalog # 12-303, Jurkat lysate. Add 2.5 µL of 2-mercaptoethanol per 100 µL of lysate and boil for 5 minutes to reduce the preparation. Use 20 µg per lane for minigels.

Application

Anti-GSK3 Antibody, clone 4G-1E detects level of GSK3 & has been published & validated for use in IC & WB.

Research Sub CategoryPI3K, Akt, & mTOR Signaling

Research CategorySignaling

Immunoprecipitation:
Not recommended.

Immunocytochemistry:
10 µg/mL of a previous lot showed positive immunostaining for GSK3 in L6 cells fixed with acetone.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General 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 51 kDa GSK-3a and a 47 kDa 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 R16;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.

Immunogen

18 residue synthetic peptide (C-KQLLHGEP NVSYICSRY) corresponding to amino acids 203-219 derived from a region of the catalytic domain (between subdomain VII and VIII) of the Drosophila GSK3/shaggy enzyme, coupled to ovalbumin. Clone 4G-1E.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Purified

Format: Purified

Protein G purified mouse IgG1 in storage buffer containing 0.02 M phosphate buffer, pH 7.6, 0.25 M NaCl, and 0.1% sodium azide.

Quality

Routinely evaluated by immunoblot on human Jurkat cells, human A431, rat L6, rat PC12 or mouse 3T3 cells.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected GSK3 in RIPA lysates from human Jurkat cells; previously GSK3 was detected in RIPA lysates from human A431, rat L6 and PC12, and mouse 3T3 cells.

Specificity

Because of the extremely high conservation of this immunogen among the known enzymes of many different species, this antibody is predicted to cross-react with nearly all species homologues.

GSK3, α and β isoforms

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

51 kDa (α) or 46 kDa (β)