Caspase-9 Rabbit Polyclonal Antibody

• Catalog Number: orb10242
• 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
• Reactivity: Canine, Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Caspase-9 subunit p35 (11-120/416aa)
• 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=1μg/test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 35/50 kDa
• Target: CASP9
• UniProt ID: P55211
• RRID: AB_10753059
• 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.

References

Chen, Liwen et al. Combination of gemcitabine and erlotinib inhibits recurrent pancreatic cancer growth in mice via the JAK-STAT pathway Oncol Rep, 39, 1081-1089 (2018)

PubMed: 29328487

Blank control: K562 (blue). Primary Antibody: Rabbit Anti-caspase-9 antibody (orb10242, 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. Protocol, The cells were fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min. Primary antibody (orb10242, 1 µg/1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (30 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min at room temperature. Acquisition of 20000 events was performed.

Blank control: RSC96 (blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice. Isotype Control Antibody: Rabbit IgG (orange), Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA, Primary Antibody Dilution: 1 µg in 100 µL 1X PBS containing 0.5% BSA (green).

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 (orb10242) 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.

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 (Insulin like growth factor 1) Polyclonal Antibody, Unconjugated at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 minutes and 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 (orb10242) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: 293T-UV Cell (Human) Lysate at 30 ug, Primary: Anti-Caspase-9 at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 35/50 kD, Observed band size: 35/50 kD.

Sample: Lane 1: Mouse Stomach tissue lysates, Lane 2: Mouse Spleen tissue lysates, Primary: Anti-Caspase-9 (orb10242) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 35/50 kDa, Observed band size: 52 kDa.

Tissue/cell: Hela 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 (orb10242) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Tissue/cell: 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 (orb10242) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Tissue/cell: human colon carcinoma, 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℃ for 20 min, Incubation: Anti-Caspase-9 Polyclonal Antibody, Unconjugated (orb10242) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/cell: MCF-7 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 (orb10242) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868589) at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.