Anti-Phospho-FOXO3a (Ser253) Polyclonal Antibody is a Rabbit antibody targeting Phospho-FOXO3a (Ser253). Anti-Phospho-FOXO3a (Ser253) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Horse,Sheep)

1. 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 (TMAB-01414) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
2. Blank control: THP-1. Primary Antibody (green line): Rabbit Anti-Phospho-FoxO3a (Ser253) antibody (TMAB-01414)
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.

IHC-P,IHC-Fr,IF,FCM

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human FoxO3a around the phosphorylation site of Ser253

Human

Forkhead Box,FOXO