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

IgG

Human,Rat (predicted:Mouse,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: 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

FOXO3

Forkhead box protein O3

Forkhead Box,FOXO

Transcriptional activator which triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. Recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3'. Participates in post-transcriptional regulation of MYC: following phosphorylation by MAPKAPK5, promotes induction of miR-34b and miR-34c expression, 2 post-transcriptional regulators of MYC that bind to the 3'UTR of MYC transcript and prevent its translation.