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Figure Legend: Up-regulation and Antibody-Peptide Competition. Peptide Competition. 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 phosphatase (5), and then incubated with the GSK-3a[pY279]/ß[pY216] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following its prior incubation with: the phosphopeptide immunogen (1), no peptide (2), the non-phosphopeptide 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 (Cat. no. ALI4404), and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to GSK-3a[pY279]/ß[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. (Cat. No. 44604G)
|Tested species reactivity||Rat , Human , Mouse|
|Published species reactivity||Rat , Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human GSK-3a/b that contains tyrosine 279/216. The sequence is conserved in rat and zebrafish GSK-3a and mouse, rat, frog and zebrafish GSK-3b.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 4 publications below|
Glycogen synthase kinase-3 beta (GSK3 beta) belongs to the subfamily of glycogen synthase kinases, that play a central role in energy metabolism, neuronal cell development, and body pattern formation. GSK3 has alpha and beta isoforms (51 kDa and 47 kDa respectively), that are functionally dissimilar. GSK3 beta plays a role in insulin and Wnt signaling, and is inhibited following the extracellular signaling events.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
DISC1 regulates expression of the neurotrophin VGF through the PI3K/AKT/CREB pathway.
44-604G was used in western blot to characterize the routes involving AKT and CREB through which DISC alters VGF expression.
|Rodríguez-Seoane C,Ramos A,Korth C,Requena JR||Journal of neurochemistry (135:598)||2015|
Terminal hypothermic Tau.P301L mice have increased Tau phosphorylation independently of glycogen synthase kinase 3α/β.
44-604G was used in western blot to study the lack of involvementof GSK3-alpha/beta in the elevated tau phosphorylation observed in Tau.P30L hypothermic mice
|Maurin H,Lechat B,Borghgraef P,Devijver H,Jaworski T,Van Leuven F||The European journal of neuroscience (40:2442)||2014|
Neurological characterization of mice deficient in GSK3α highlight pleiotropic physiological functions in cognition and pathological activity as Tau kinase.
44-604G was used in western blot to investigate the contribution of GSK3α to neurological diseases.
|Maurin H,Lechat B,Dewachter I,Ris L,Louis JV,Borghgraef P,Devijver H,Jaworski T,Van Leuven F||Molecular brain (6:null)||2013|
Regulation and localization of tyrosine216 phosphorylation of glycogen synthase kinase-3beta in cellular and animal models of neuronal degeneration.
||Bhat RV,Shanley J,Correll MP,Fieles WE,Keith RA,Scott CW,Lee CM||Proceedings of the National Academy of Sciences of the United States of America (97:11074)||2000|
FA, GSK-3 beta, factor A, glycogen synthase kinase-3 beta, serine/threonine-protein kinase GSK3B, GSK3beta isoform, Glycogen Synthase Kinase 3 beta, EC 184.108.40.206, GSK-3 beta Factor A, GSK3B
GSK-3beta, GSK3, 7330414F15Rik, GSK-3, C86142, 8430431H08Rik
Carbohydrate metabolism Other polysaccharide metabolism Glycogen metabolism Protein metabolism and modification Protein modification Protein phosphorylation Signal transduction Cell surface receptor mediated signal transduction Other receptor mediated signaling pathway Intracellular signaling cascade Developmental processes Embryogenesis Segment specification Ectoderm development Neurogenesis Mesoderm development Cell cycle Mitosis Cell proliferation and differentiation