Phospho-FoxO3a (Ser253) Rabbit Polyclonal Antibody

• Catalog Number: orb6053
• Category: Antibodies
• Description: Phospho-FoxO3a (Ser253) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Porcine, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human FoxO3a around the phosphorylation site of Ser253 AV(p-S)MD
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 71 kDa
• Target: FOXO3
• UniProt ID: O43524
• RRID: AB_10928217
• 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: FOXO3A(phospho S253); p=FOXO3A(phospho S253); FOXO
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: THP-1. Primary Antibody (green line): Rabbit Anti-Phospho-FoxO3a (Ser253) antibody (orb6053), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 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 -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 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.

Paraformaldehyde-fixed, paraffin embedded (Rat brain), Antigen retrieval by microwave in sodium citrate buffer (pH6.0), Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes, Blocking buffer (3% BSA) at RT for 30 min, Antibody incubation with (Phospho-FoxO3a (Ser253)) Polyclonal Antibody, Unconjugated (orb6053) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.