Phospho-Caspase-9 (Thr125) Rabbit Polyclonal Antibody

• Catalog Number: orb5767
• Category: Antibodies
• Description: Phospho-Caspase-9 (Thr125) 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 Caspase-9 around the phosphorylation site of Thr125 PE(p-T)PR
• 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=0.2μg /test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 35 kDa
• Target: CASP9
• UniProt ID: P55211
• RRID: AB_10927991
• 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: APAF 3; APAF3; Apoptosis related cysteine peptidas
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody (green line): Rabbit Anti-Phospho-Caspase-9 (Thr125) antibody (orb5767), dilution: 0.2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 1 µg/Test.

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-Caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (orb5767) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat 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 (P-caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (orb5767) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat 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 (P-caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (orb5767) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Brain (Rat) Lysate at 40 ug, Primary: Anti-p-Caspase-9 (Thr125) (orb5767) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution, Predicted band size: 35kD, Observed band size: 35kD.

Sample: H9C2 (Rat) cell Lysate at 40 ug, Primary: Anti-p-Caspase-9 (Thr125) (orb5767) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution, Predicted band size: 35kD, Observed band size: 35kD.

Sample: Heart (Rat) Lysate at 40 ug, Heart (Mouse) Lysate at 40 ug, Primary: Anti-Phospho-Caspase-9 (Thr125) (orb5767) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46-51/35/37 kD, Observed band size: 51/37/35 kD.