bsm-54505R
[Primary Antibody]
Phospho-GSK3(alpha+beta)(Y216+Y279) Monoclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: Phospho-GSK3(alpha+beta)(Y216+Y279)
Modification Site: Tyr216+Tyr279
Clonality: Monoclonal
Isotype: IgG
Entrez Gene: 2932
Swiss Prot: P49840
Source: Synthetic phospho-Peptide corresponding to residues surrounding Tyr216 and 279 of human GSK3(alpha+beta).
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Background:
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. GSK3s substrate specificity is unique in that phosphorylation of substrate only occurs if a phosphoserine or phosphotyrosine is present four residues C terminal to the site of GSK phosphorylation. 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. Phosphorylation of GSK3 beta on Ser9 (Ser21 in GSK3 alpha) by protein kinase B (PKB) causes its inactivation is the primary mechanism responsible for growth factor inhibition of this kinase. 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. GSK3 beta also has been shown to phosphorylate aberrant sites on the microtubule associated protein tau, which is critical for the progression of Alzheimer's disease. GSK3B is involved in energy metabolism, neuronal cell development, and body pattern formation.
Size: 100ul
Concentration: 1ug/ul
Predicted Molecular Weight: 51
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Phospho-GSK3(alpha+beta)(Y216+Y279) Monoclonal Antibody, Unconjugated (bsm-54505R) at 1:50 for 30 minutes at room temperature, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Phospho-GSK3(alpha+beta)(Y216+Y279) Monoclonal Antibody, Unconjugated (bsm-54505R) at 1:50 for 30 minutes at room temperature, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Phospho-GSK3(alpha+beta)(Y216+Y279) Monoclonal Antibody, Unconjugated (bsm-54505R) at 1:50 for 30 minutes at room temperature, DAB staining.
A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-GSK3(alpha+beta)(Y216+Y279)) Monoclonal Antibody, Unconjugated (bsm-54505R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-GSK3(alpha+beta)(Y216+Y279)) Monoclonal Antibody, Unconjugated (bsm-54505R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-GSK3(alpha+beta)(Y216+Y279)) Monoclonal Antibody, Unconjugated (bsm-54505R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Mouse Testis tissue lysates; Lane 3: Rat Cerebrum tissue lysates; Lane 4: Rat Testis tissue lysates; Lane 5: Human HeLa cell lysates; Lane 6: Human 293T cell lysates; Lane 7: Human HepG2 cell lysates; Lane 8: Human MCF-7 cell lysates probed with phospho-GSK3 Alpha + Beta (Tyr279+Tyr216) Monoclonal Antibody, Unconjugated (bsm-54505R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.