Anti-Phospho-Caspase-9 (Thr125) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Caspase-9 (Thr125). Anti-Phospho-Caspase-9 (Thr125) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (Phospho-Caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (TMAB-10444) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (P-caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (TMAB-10444) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (P-caspase-9 (Thr125)) Polyclonal Antibody, Unconjugated (TMAB-10444) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
4. Sample: H9C2 (Rat) Cell Lysate at 40 μg
Primary: Anti-p-Caspase-9 (Thr125) (TMAB-10444) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 35 kD
5. Sample: Brain (Rat) Lysate at 40 μg
Primary: Anti-p-Caspase-9 (Thr125) (TMAB-10444) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 35 kD
6. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-Phospho-Caspase-9 (Thr125) antibody (TMAB-10444), Dilution: 0.2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg /test.
7. Sample:
Heart (Rat) Lysate at 40 μg
Heart (Mouse) Lysate at 40 μg
Primary: Anti-Phospho-Caspase-9 (Thr125) (TMAB-10444) 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: 51/37/35 kD

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 0.2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Caspase-9 around the phosphorylation site of Thr125

Human

CASP9

Caspase-9

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.