Rabbit anti-Caspase-9 Polyclonal Antibody

Blank control: K562 (blue). Primary Antibody:Rabbit Anti-caspase-9 antibodyGreen); Dilution: 1ug in 100 uL 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 1ug/1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA+10% goat serum (30min) 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 20,000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (rat heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated at 1:200 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 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) instructionsand DAB staining.

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 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.

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