Anti-Phospho-Bcl2 (Ser87) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-Bcl2 (Ser87). Anti-Phospho-Bcl2 (Ser87) Polyclonal Antibody 2 can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Rat (predicted:Mouse,Horse,Rabbit)

1. Paraformaldehyde-fixed, paraffin embedded (rat lung); 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 (BCL2) Polyclonal Antibody, Unconjugated (TMAB-10378) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
2. Blank control (blue): K562 (fixed with 2% paraformaldehyde for 10 min at 37℃). Primary Antibody: Rabbit Anti-Phospho-Bcl2 (Ser87) antibody (TMAB-10378, Green); Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions; Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue), Dilution: 1: 200 in 1 X PBS containing 0.5% BSA.
3. Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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-Bcl2 (Ser87)) Polyclonal Antibody, Unconjugated (TMAB-10378) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (rat spleen); 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-Bcl2 (Ser87)) Polyclonal Antibody, Unconjugated (TMAB-10378) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (rat lymphoid); 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-Bcl2 (Ser87)) Polyclonal Antibody, Unconjugated (TMAB-10378) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Blank control: HL-60.
Primary Antibody (green line): Rabbit Anti-Phospho-Bcl2 (Ser87) antibody (TMAB-10378)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1%PBST for 20 min at room temperature. 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 Bcl2 around the phosphorylation site of Ser87

Human

BCL2

Apoptosis regulator Bcl-2

Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1).