Phospho-Bcl2 (Thr69) Rabbit Polyclonal Antibody

• Catalog Number: orb155808
• Category: Antibodies
• Description: Phospho-Bcl2 (Thr69) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human Bcl-2 around the phosphorylation site of Thr69 AR(p-T)SP
• Antibody Type: Primary Antibody
• 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
• 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: Bcl2(phospho-Thr69); Bcl2(phospho-T69); Bcl2(phosp
• 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-Bcl2 (Thr69) antibody (orb155808, 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-Bcl2 (Thr69) antibody (orb155808), 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 (rat stomach 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-Bcl2 (Thr69)) Polyclonal Antibody, Unconjugated (orb155808) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Spleen (Mouse) Lysate at 40 ug, LOVO Cell (Human) Lysate at 40 ug, HL60 Cell (Human) Lysate at 40 ug, U937 Cell (Human) Lysate at 40 ug, MCF-7 Cell (Human) Lysate at 40 ug, Hela Cell (Human) Lysate at 40 ug, Primary: Anti-Phospho-Bcl2 (Thr69) (orb155808) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 26 kD, Observed band size: 26 kD.

Tissue/Cell: rat lung tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, 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°C for 20 min, Incubation: Anti-Phospho-Bcl2 (Thr69) Polyclonal Antibody, Unconjugated (orb155808) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.