Anti-Phospho-BNIP3 (Ser95) Polyclonal Antibody is a Rabbit antibody targeting Phospho-BNIP3 (Ser95). Anti-Phospho-BNIP3 (Ser95) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human (predicted:Dog,Cow,Horse,Sheep)

1. Tissue/cell: human liver carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, 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℃ for 20 min;
Incubation: Anti-phospho-BNIP3 (Ser95) polyclonal Antibody, Unconjugated (TMAB-10408) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Paraformaldehyde-fixed, paraffin embedded (Human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-BNIP3 (Ser95)) Polyclonal Antibody, Unconjugated (TMAB-10408) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-phospho-BNIP3 (Ser95) antibody (TMAB-10408)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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℃, 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 20,000 events was performed.

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

Polyclonal

KLH conjugated synthesised phosphopeptide: human BNIP3 around the phosphorylation site of Ser95

Human

BNIP3

BCL2/adenovirus E1B 19 kDa protein-interacting protein 3

Apoptosis-inducing protein that, which can overcome BCL2 suppression. May play a role in repartitioning calcium between the two major intracellular calcium stores in association with BCL2.