Anti-phospho-GSK3 (Tyr279/216)

Anti-phospho-GSK3 (Tyr279/Tyr216), clone 5G-2F

Glycogen synthase kinase 3 (GSK3) is a proline directed serine threonine kinase that was initially identified as a phosphorylating and inactivating glycogen synthase, a key enzyme in glycogen metabolism. Since then, it has been shown to be involved in the regulation of a diverse array of cellular functions, including protein synthesis, cell proliferation, cell differentiation, microtubule assembly/disassembly, and apoptosis.
There exists two isoforms of GSK3, alpha and beta, and they show a high degree of amino acid homology. The two isoforms of GSK3 are strictly regulated via phosphorylation. Activation of GSK3 beta is dependent upon the phosphorylation of Tyr216 (Tyr279 in GSK3 alpha). Upon activation, it has been shown to phosphorylate a number of different cellular proteins, including p53, c-Myc, c-Jun, heat shock factor 1 (HSF1), and cyclin D1

• GSK-3 alpha
• glycogen synthase kinase 3 alpha

5G-2F

This antibody recognizes GSK3 is constitutively phosphorylated at this Tyrosine residue in resting cells. This immunogen is identical in 15 of 17 residues in human and rat GSK3α (Tyr279) and GSK3β (Tyr216). The phosphotyrosine residue can be removed from GSK3 by treatment with PTP1B.

Western Blotting

Immunoprecipitation

• Human
• Rat
• Mouse
• Dictyostelium
• Yeast (S. cerevisiae )

Mouse, rat, human, S. cerevisiae and Dictyostelium. Clone 5G-2F should cross-react with nearly all species because of the extremely high conservation of this region among the known enzymes of many different species.

IgG1κ

Synthetic peptide (C-KQLLHGEPNVS [pY]ICSRY) 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.

a.a. 203-219

Phosphorylation

46 kDa (alpha) and 51 kDa (beta)

Jurkat cell lysate, or serum starve 3T3-L1 cells for 3 hours, then stimulate with 100 nM insulin or 100 ng/mL IGF 1 for 5 min.
Included Positive Antigen Control: Catalog # 12-303, Jurkat lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Frozen at -20ºC,

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 -20ºC. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

• GSK3A
• DKFZp686D0638
• EC 2.7.11.26

Routinely evaluated by immunoblot on human Jurkat cells, mouse 3T3/A31, human A431, or rat L6 cells.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected phospho-GSK3 in RIPA lysates from human Jurkat cells. Previous lots detected phospho-GSK3 in mouse 3T3/A31, human A431 lysates and rat L6 cells

Signaling

PI3K, Akt, & mTOR Signaling

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.

Upstate (Millipore)

1 mg/mL

Monoclonal Antibody

Purified

Mouse

Protein G chromatography