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||Human, Mouse, Rat|
|Published species reactivity||Rat, Human, Mouse, Not Applicable|
|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.
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.
The identification of raft-derived tau-associated vesicles that are incorporated into immature tangles and paired helical filaments.
44-604G was used in immunohistochemistry - paraffin section to study the relationship between neurofibrillary tangles and raft domains
|Nishikawa T,Takahashi T,Nakamori M,Hosomi N,Maruyama H,Miyazaki Y,Izumi Y,Matsumoto M||Neuropathology and applied neurobiology (42:639)||2016|
A reverse-phase protein microarray-based screen identifies host signaling dynamics upon Burkholderia spp. infection.
44-604G was used to identify host signaling dynamics upon Burkholderia spp. infection by a reverse-phase protein microarray-based screen
|Chiang CY,Uzoma I,Lane DJ,Memi¿evi¿ V,Alem F,Yao K,Kota KP,Bavari S,Wallqvist A,Hakami RM,Panchal RG||Frontiers in microbiology (6:null)||2015|
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 3¿/ß.
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 GSK3¿ 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|
|Developmental regulation of tau phosphorylation, tau kinases, and tau phosphatases.||Yu Y,Run X,Liang Z,Li Y,Liu F,Liu Y,Iqbal K,Grundke-Iqbal I,Gong CX||Journal of neurochemistry (108:1480)||2009|
|Not Applicable||Not Cited||
Measuring GSK3 expression and activity in cells.
44-604G was used in western blot to discuss the benefits and limitations of different assays to measure glycogen synthase kinase activity
|Cole AR,Sutherland C||Methods in molecular biology (Clifton, N.J.) (468:45)||2008|
|Not Applicable||Not Cited||
Interaction between ERK and GSK3beta mediates basic fibroblast growth factor-induced apoptosis in SK-N-MC neuroblastoma cells.
44-604G was used in western blot to elucidate how treatment with basic fibroblast growth factor suppresses the growth of Ewing's sarcoma family of tumors
|Ma C,Bower KA,Chen G,Shi X,Ke ZJ,Luo J||The Journal of biological chemistry (283:9248)||2008|
Calcium-mediated transient phosphorylation of tau and amyloid precursor protein followed by intraneuronal amyloid-beta accumulation.
44-604G was used in western blot to investigate how cytosolic calcium alters neuronal metabolism of amyloid precursor protein
|Pierrot N,Santos SF,Feyt C,Morel M,Brion JP,Octave JN||The Journal of biological chemistry (281:39907)||2006|
Effect of aged garlic extract on APP processing and tau phosphorylation in Alzheimer's transgenic model Tg2576.
44-604G was used in western blot to compare the anti-amyloidogenic, anti-inflammatory, and anti-tangle effects of dietary aged garlic extract with its prominent constituents in a mouse model of Alzheimer's disease
|Chauhan NB||Journal of ethnopharmacology (108:385)||2006|
|Rat||Not Cited||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|
Tauopathy contributes to synaptic and cognitive deficits in a murine model for Alzheimer's disease.
44-604G was used in immunohistochemistry (free floating) to present data that suggests that pathological Aβ species induce changes in Tau that contribute to cognitive deficits correlating with synaptic deficits and hippocampal atrophy in an Alzheimer's disease model.
|Stancu IC,Ris L,Vasconcelos B,Marinangeli C,Goeminne L,Laporte V,Haylani LE,Couturier J,Schakman O,Gailly P,Pierrot N,Kienlen-Campard P,Octave JN,Dewachter I||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (28:2620)||2014|