Anti-APAF1(NT) Polyclonal Antibody is a Rabbit antibody targeting APAF1(NT). Anti-APAF1(NT) Polyclonal Antibody can be used in FCM,ICC/IF,IF,IHC-Fr,IHC-P,WB.

IgG

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

1. Tissue/cell: rat lung tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), 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-APAF1 (NT) Polyclonal Antibody, Unconjugated (TMAB-00126) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), 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-APAF1 (NT) Polyclonal Antibody, Unconjugated (TMAB-00126) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Sample:
Lane 1: Lung (Mouse) Lysate at 40 μg
Lane 2: Cerebrum (Mouse) Lysate at 40 μg
Primary: Anti-APAF1 (NT) (TMAB-00126) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 140 kDa
Observed band size: 140 kDa
4. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (APAF1 (NT)) Polyclonal Antibody, Unconjugated (TMAB-00126) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (APAF1 (NT)) Polyclonal Antibody, Unconjugated (TMAB-00126) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Tissue/cell: SH-SY5Y 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 (APAF1 (NT)) polyclonal Antibody, Unconjugated (TMAB-00126) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
7. Tissue/cell: SH-SY5Y 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 (APAF1 (NT)) polyclonal Antibody, Unconjugated (TMAB-00126) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
8. U-937 cells were fixed with 4% PFA for 10 min at room temperature,permeabilized with 20% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with APAF1 (NT) Antibody (TMAB-00126) at 1:500 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 0.2ug/test

Polyclonal

KLH conjugated synthetic peptide: human Apaf-1

Human

APAF1

Apoptotic protease-activating factor 1

Cytochrome C,Metabolism,Mitochondrial,Cytochrome C,Mitochondrial,Apoptosis,Mitochondrial markers

Oligomeric Apaf-1 mediates the cytochrome c-dependent autocatalytic activation of pro-caspase-9 (Apaf-3), leading to the activation of caspase-3 and apoptosis. This activation requires ATP. Isoform 6 is less effective in inducing apoptosis.