Phospho-GSK-3 Beta (Ser21) Rabbit Polyclonal Antibody

• Catalog Number: orb5357
• Category: Antibodies
• Description: Phospho-GSK-3 Beta (Ser21) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Guinea pig, Porcine, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human GSK-3 Beta around the phosphorylation site of Ser21 QP(p-S)AF
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=3μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 47 kDa
• Target: GSK3B
• UniProt ID: P49841
• RRID: AB_10924194
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: GSK3B(phospho-Ser21); GSK3B(phospho-S21); p-GSK-3
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (Black line): Mouse spleen (Black). Primary Antibody (green line): Rabbit Anti-GSK-3 Beta (Ser21) antibody (orb5357), dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5% BSA goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Sample: Brain (Mouse) Lysate at 40 ug, Cerebellum (Mouse) Lysate at 40 ug, Primary: Anti-phospho-GSK-3 Beta (Ser21) (orb5357) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 47 kD, Observed band size: 49 kD.

Sample: Lane 1: Muscle (Mouse) Lysate at 40 ug, Lane 2: Liver (Mouse) Lysate at 40 ug, Lane 3: Cerebrum (Mouse) Lysate at 40 ug, Lane 4: Pancreas (Mouse) Lysate at 40 ug, Lane 5: Muscle (Rat) Lysate at 40 ug, Lane 6: Cerebrum (Rat) Lysate at 40 ug, Lane 7: Pancreas (Rat) Lysate at 40 ug, Primary: Anti-phospho-GSK-3 Beta (Ser21) (orb5357) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 47 kD, Observed band size: 47 kD.

Tissue/Cell: rat testis tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-phospho-GSK-3 Beta (Ser21) Polyclonal Antibody, Unconjugated (orb5357) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.