Anti-caspase-9 p10 Polyclonal Antibody is a Rabbit antibody targeting caspase-9 p10. Anti-caspase-9 p10 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Horse,Rabbit)

1. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-caspase-9 p10 Polyclonal Antibody, Unconjugated (TMAB-03674) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Blank control: K562 (fixed with 80% methanol (5 min) and and then permeabilized with 0.01 M Pcaspase-9 p10 antibody (TMAB-03674, 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.
3. 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 30 min; Antibody incubation with (caspase-9 p10) Polyclonal Antibody, Unconjugated (TMAB-03674) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. 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 (TMAB-03674) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 10/46 kD
Observed band size: 46 kD
5. Blank control: K562.
Primary Antibody (green line): Rabbit Anti-caspase-9 p10 antibody (TMAB-03674)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5μg /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℃. 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 20,000 events was performed.

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

Polyclonal

KLH conjugated synthetic peptide: human caspase-9 subunit p10

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.