General description

Glycogen synthase kinase-3 beta/alpha (UniProts: P49841 and P49840; also known as EC: 2.7.11.26 and 2.7.11.26, GSK-3 beta/alpha, Serine/threonine-protein kinase GSK3B/alpha) is encoded by the GSK3B/GSK3A gene (Gene ID: 2932/2931) in human. GSK-3 is a multifunctional serine/threonine kinase, is a key regulator of numerous signaling pathways during embryogenesis and in metabolic control. Two isoforms of GSK-3 are reported in mammals: a 51 kDa GSK-3 alpha and a 47 kDa GSK-3 beta. The GSK-3 alpha contains a glycine-rich extension at its N-terminal. GSK-3 is constitutively active in cells and is regulated through inhibition of its activity. GSK-3 shows a preference for target proteins that are pre-phosphorylated at a `priming residue located C-terminal to the site of GSK-3 phosphorylation. Priming phosphorylation, although not absolutely required, enhances the efficiency of phosphorylation of most GSK-3 substrates. Phosphorylation of a threonine residue in the activation loop (T-loop) is considered to be essential for its activity. The T-loop of GSK-3 alpha is phosphorylated at Tyr279 and GSK-3 beta at Tyr216, which play a role in forcing open the substrate-binding site of the enzyme. A primary mechanism of GSK3 activity regulation is phosphorylation of N-terminal serine residues (Serine 9 in GSK3 beta and serine 21 in GSK3 alpha). Phosphorylation of these sites is inhibitory to GSK3 kinase activity because the phosphorylated N-terminus acts as a competitive inhibitor for primed substrates. GSK-3 beta is constitutively active in resting cells and treatment of cells with an agent, such as insulin, causes GSK-3 inactivation through a PI 3-kinase (PI 3-K)-dependent mechanism.

Specificity

Clone 15C2 detects non-phosphorylated forms of both GSK-3 beta and GSK-3 alpha in Human, Mouse, and Rat. It targets an epitope within the N-terminal region.

Immunogen

Epitope: N-terminus

KLH-conjugated linear peptide corresponding to 14 amino acids from the N-terminal region of human GSK-3 beta.

Application

Anti-nonphospho-Ser9/21 GSK3 beta/alpha, clone 15C2, Cat. No. MABN2465, is a highly specific mouse monoclonal antibody that targets nonphospho Ser9/21 GSK3 beta/alpha and has been testedfor use in Immunocytochemistry, Immunohistochemistry (Paraffin), Immunoprecipitation and Western Blotting.

Research Category
Neuroscience

Western Blotting Analysis: A 1:500 dilution from a representative lot detected nonphospho-Ser9/21 GSK3 / in 10 µg of human brain tissue lysate.

Immunohistochemistry Analysis: A 1:50 dilution from a representative lot detected nonphospho-Ser9/21 GSK3 / in human cerebral cortex and rat cerebral cortex tissues.

Immunoprecipitation Analysis: A representative lot detected nonphospho-Ser9/21 GSK3 / in Immunoprecipitation applications (Grabinski, T., et. al. (2016). Front Mol Neurosci. 9:123).

Western Blotting Analysis: A representative lot detected nonphospho-Ser9/21 GSK3 / in Western Blotting applications (Grabinski, T., et. al. (2016). Front Mol Neurosci. 9:123).

Immunocytochemistry Analysis: A representative lot detected nonphospho-Ser9/21 GSK3 / in Immunocytochemistry applications (Grabinski, T., et. al. (2016). Front Mol Neurosci. 9:123).

Immunohistochemistry Analysis: A representative lot detected nonphospho-Ser9/21 GSK3 / in Immunohistochemistry applications (Grabinski, T., et. al. (2016). Front Mol Neurosci. 9:123).

Quality

Evaluated by Western Blotting in rat brain tissue lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected nonphospho-Ser9/21 GSK3 beta/alpha in 10 µg of rat brain tissue lysate.

Target description

46.74/50.98 kDa calculated for GSK3 beta and alpha, respectively. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

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.