Caspase-9 Rabbit Polyclonal Antibody

• Catalog Number: orb500803
• Category: Antibodies
• Description: Caspase-9 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Caspase-9 (231-330/416aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 35/50 kDa
• Target: CASP9
• UniProt ID: P55211
• 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: Caspase-9 subunit p35; Apaf-3; APAF 3; APAF3; Apop
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

HepG2 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Caspase-9) polyclonal Antibody, Unconjugated (orb500803) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Overlay histogram showing Hela cells (1 ug/1x10^6 cells) stained with orb500803 (Green line). The cells were fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min. The cells were then incubated in 1x PBS/10% normal goat serum to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was fluorescein isothiocyanate goat anti-rabbit IgG (H+L) (orb868805, Brillant blue line) at 1/200 dilution for 30 min at 22°C. Isotype control antibody was rabbit IgG (polyclonal, Orange line) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of 20000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Paraformaldehyde-fixed, paraffin embedded (mouse pancreas), 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 (Caspase-9) Polyclonal Antibody, Unconjugated (orb500803) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat heart), 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 (Caspase-9) Polyclonal Antibody, Unconjugated (orb500803) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat pancreas), 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 (Caspase-9) Polyclonal Antibody, Unconjugated (orb500803) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat urinary bladder), 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 (Caspase-9) Polyclonal Antibody, Unconjugated (orb500803) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: K562 Cell (Human) Lysate at 40 ug, Primary: Anti-Caspase-9 (orb500803) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 35/50 kD, Observed band size: 35 kD.

Sample: Spleen (Mouse) Lysate at 40 ug, Primary: Anti-Caspase-9 (orb500803) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 35/50 kD, Observed band size: 35 kD.

Sample: Urinary bladder (Mouse) Lysate at 40 ug, Jurkat (Human) Cell Lysate at 30 ug, Primary: Anti-Caspase-9 (orb500803) 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: 35 kD.