Caspase-9 p10 Rabbit Polyclonal Antibody

• Catalog Number: orb1677
• Category: Antibodies
• Description: Caspase-9 p10 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human caspase-9 subunit p10 (351-416/416aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:50-200, Flow-Cyt=1μg /Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 10/50 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: APAF 3; APAF3; Apoptosis related cysteine peptidas
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Sample: Brain (Mouse) Lysate at 40 ug, Lung (Mouse) Lysate at 40 ug, Primary: Anti-caspase-9 p10 (orb1677) at 1/300 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1/5000 dilution, Predicted band size: 10/50 kD, Observed band size: 50 kD.

Blank control: K562 (fixed with 80% methanol (5 min) and and then permeabilized with 0.01M PBS-Tween for 20 min). Primary Antibody: Rabbit Anti-caspase-9 p10 antibody (orb1677, 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: K562. Primary Antibody (green line): Rabbit Anti-caspase-9 p10 antibody (orb1677), dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. 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 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 p10) Polyclonal Antibody, Unconjugated (orb1677) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Jurkat (Human) Cell Lysate at 30 ug, Hela (Human) Cell Lysate at 30 ug, HepG2 (Human) Cell Lysate at 30 ug, Primary: Anti-caspase-9 p10 (orb1677) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46-51/37/35/10 kD, Observed band size: 46/37 kD.

Sample: Lane 1: Esophagus (Mouse) Lysate at 40 ug, Lane 2: Stomach (Mouse) Lysate at 40 ug, Lane 3: Urinary bladder (Mouse) Lysate at 40 ug, Primary: Anti-caspase-9 p10 (orb1677) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46-51/37/35/10 kD, Observed band size: 46 kD.

Tissue/Cell: rat brain 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-caspase-9 p10 Polyclonal Antibody, Unconjugated (orb1677) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.