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Synthetic peptide derived from the regions of GSK 3 alpha + beta protein that contain tyrosine 216/279.
Glycogen synthase kinase-3 (GSK 3) is a proline-directed serine-threonine kinase that was initially identified as a phosphorylating and inactivating glycogen synthase. Two isoforms, alpha (GSK 3A) and beta, show a high degree of amino acid homology. GSK 3B is involved in energy metabolism, neuronal cell development, and body pattern formation.
Our Abpromise guarantee covers the use of ab4797 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 51,47 kDa. Use at a concentration of 0.5 - 1.0 µg/ml. Predicted molecular weight: 51kDa for GSK 3 alpha and 47 kDa for GSK 3 beta.|
|Dot Blot||Use at an assay dependent dilution.|
|ELISA||Use at an assay dependent dilution.|
Extracts of 3T3L1 cells stimulated with 100 nM insulin for 10 minutes were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature and either left untreated (1-4) or treated with lambda (λ) phosphatase (5), then incubated with the GSK-3 α [pY279] / β [pY216] antibody for two hours at room temperature in a 1% BSATBST buffer, following its prior incubation with: the phosphopeptide immunogen (1), no peptide (2), the non-hosphopeptide corresponding to the phosphopeptide immunogen (3), or a generic phosphotyrosine-containing peptide (4). After washing, the membrane was incubated with goat F(ab’) 2 anti-rabbit IgG HRP conjugate, and signals were detected. The data show that only the phosphopeptide corresponding to GSK-3α [pY279] /β[pY216] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, further verifying that the antibody is phospho-specific.
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