Anti-Caspase-9 Polyclonal Antibody 3 is a Rabbit antibody targeting Caspase-9. Anti-Caspase-9 Polyclonal Antibody 3 can be used in FCM,ICC/IF,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Mouse,Rat (predicted:Pig,Cow,Rabbit,Sheep)

1. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (TMAB-00294) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (TMAB-00294) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Sample:
Spleen (Mouse) Lysate at 40 μg
Primary: Anti-Caspase-9 (TMAB-00294) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35/50 kDa
Observed band size: 35 kDa
4. Sample:
Urinary bladder (Mouse) Lysate at 40 μg
Jurkat (Human) Cell Lysate at 30 μg
Primary: Anti-Caspase-9 (TMAB-00294) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46-51/35/37 kDa
Observed band size: 35 kDa
5. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (TMAB-00294) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Caspase-9) Polyclonal Antibody, Unconjugated (TMAB-00294) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. 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 (TMAB-00294) 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 nucleus.
8. Sample: K562 Cell (Human) Lysate at 40 μg
Primary: Anti-Caspase-9 (TMAB-00294) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35/50 kDa
Observed band size: 35 kDa
9. Overlay histogram showing Hela cells (1 μg/1x10^6 cells) stained with TMAB-00294 (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) 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 20,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated synthetic peptide: human Caspase-9

Human

CASP9

Caspase-9

Caspases,Cytochrome C,Metabolism,Caspases,Cytochrome C,Caspases,Apoptosis

Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).
Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9.