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05-413

Sigma-Aldrich

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

clone 5G-2F, Upstate®, from mouse

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Synonym(s):
GSK-3 alpha, glycogen synthase kinase 3 alpha
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

5G-2F, monoclonal

species reactivity

mouse, human, rat

manufacturer/tradename

Upstate®

technique(s)

western blot: suitable

isotype

IgG1κ

suitability

not suitable for immunoprecipitation

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

phosphorylation (pTyr279/pTyr216)

Gene Information

human ... GSK3A(2931)
mouse ... Gsk3A(606496)
rat ... Gsk3A(50686)

General description

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

Specificity

Predicted to react with GSK-3 orthologs from drosophila melanogaster and many other species based on sequence homology surrounding the target phosphotyrosine residue.
Recognizes GSK-3 phosphorylated at the target tyrosine residue (human GSK-3α pY279 and GSK-3β pY216). The 17-amino acid immunogen peptide sequence differs from the corresponding human, mouse, and rat sequence by only two amino acid residues.

Immunogen

Epitope: Surrounding pTyr214 (Drosophila GSK-3), pTyr279 (human GSK3α), pTyr216 (human GSK3β)
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.

Application

Research Category
Signaling
Research Sub Category
PI3K, Akt, & mTOR Signaling
This Anti-phospho-GSK3 (Tyr279/Tyr216) Antibody, clone 5G-2F is validated for use in WB for the detection of phospho-GSK3 (Tyr279/Tyr216).

Quality

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

Target description

51 kDa (alpha) and 46 kDa (beta)

Physical form

Format: Purified
Protein G Chromatography
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.

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

Analysis Note

Control
Positive Antigen Control: Catalog #12-303, Jurkat cell lysate.

Other Notes

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

Legal Information

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

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.

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Xiaohong Wang et al.
The Journal of biological chemistry, 279(1), 133-141 (2003-10-21)
To gain insight into the mechanism by which the adapter protein SH2-B promotes nerve growth factor (NGF)-mediated neuronal differentiation and survival, the effect of SH2-B on the serine/threonine kinase Akt/protein kinase B and downstream effector proteins was examined. PC12 cells
Arnold Johnson
American journal of physiology. Lung cellular and molecular physiology, 296(4), L700-L709 (2009-02-17)
The hypothesis tested was PKCalpha mediates the phosphorylation of glycogen synthetase kinase 3beta (GSK3beta) and that the GSK3beta inhibition modulates the response to tumor necrosis factor-alpha (TNF) in rat pulmonary microvessel endothelial cells (PMEC). PMEC were treated with TNF for
Insulin stimulation of glycogen synthesis and glycogen synthase activity is blocked by wortmannin and rapamycin in 3T3-L1 adipocytes: evidence for the involvement of phosphoinositide 3-kinase and p70 ribosomal protein-S6 kinase.
Shepherd, P R, et al.
The Biochemical Journal, 305 ( Pt 1), 25-28 (1995)
Baculovirus-mediated expression and characterisation of rat glycogen synthase kinase-3 beta, the mammalian homologue of the Drosophila melanogaster zeste-white 3sgg homeotic gene product.
Hughes, K, et al.
European Journal of Biochemistry, 203, 305-311 (1992)
Compression alters kinase and phosphatase activity and tau and MAP2 phosphorylation transiently while inducing the fast adaptive dendritic remodeling of underlying cortical neurons.
Li-Jin Chen,Yueh-Jan Wang,Guo-Fang Tseng
Journal of Neurotrauma null

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