Phospho-Bcl-2 (Thr129) Rabbit Polyclonal Antibody

• Catalog Number: orb4669
• Category: Antibodies
• Description: Phospho-Bcl-2 (Thr129) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Porcine, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human Bcl-2 around the phosphorylation site of Thr129 FA(p-T)VV
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 26 kDa
• Target: BCL2
• UniProt ID: P10415
• RRID: AB_10941101
• 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: Bcl-2 (phospho T129); Bcl-2 (phospho Thr129); p-Bc
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue): K562 (fixed with 2% paraformaldehyde for 10 min at 37°C). Primary Antibody: Rabbit Anti-Phospho-Bcl-2 (Thr129) antibody (orb4669, 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 1X PBS containing 0.5% BSA.

Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-Phospho-Bcl-2 (Thr129) antibody (orb4669), 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 20000 events was performed.

Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-Phospho-Bcl-2 (Thr129) antibody (orb4669), 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 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), 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-Bcl-2 (Thr129)) Polyclonal Antibody, Unconjugated (orb4669) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse 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-Bcl-2 (Thr129)) Polyclonal Antibody, Unconjugated (orb4669) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse ovary), 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-Bcl-2 (Thr129)) Polyclonal Antibody, Unconjugated (orb4669) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat ovary tissue), 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 (P-Bcl-2 (Thr129)) Polyclonal Antibody, Unconjugated (orb4669) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat ovary), 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 (orb4669) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

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-Bcl-2 (Thr129)) Polyclonal Antibody, Unconjugated (orb4669) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Heart (Mouse) Lysate at 40 ug, Primary: Anti-Phospho-Bcl-2 (Thr129) (orb4669) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 26 kD, Observed band size: 26 kD.

Sample: HL60 (Human) Cell Lysate at 40 ug, Primary: Anti-Phospho-Bcl-2 (Thr129) (orb4669) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 26 kD, Observed band size: 26 kD.