P53 (wt-p53) Rabbit Polyclonal Antibody

Blank control (blue line): Hela (blue). Primary Antibody (green line): Rabbit Anti-P53 (wt-p53) antibody (orb13631), Dilution: 1 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. Protocol, The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Sample: Liver (Mouse) Lysate at 40 ug, Liver (Rat) Lysate at 40 ug, Primary: Anti-P53 (wt-p53) (orb13631) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 53 kD, Observed band size: 53 kD.

Sample: MCF-7 (Human) Cell Lysate at 40 ug, DU145 (Human) Cell Lysate at 40 ug, Primary: Anti-P53 (wt-p53) (orb13631) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 53 kD, Observed band size: 53 kD.

Tissue/Cell: A549 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 (P53 (wt-p53)) polyclonal Antibody, Unconjugated (orb13631) 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 nuclei.

Tissue/Cell: A549 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 (P53 (wt-p53)) polyclonal Antibody, Unconjugated (orb13631) 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 nuclei.

Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), 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-P53 (wt-p53) Polyclonal Antibody, Unconjugated (orb13631) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.