Mutant p53 (N235K, N239Y) Rabb

Blank control (blue line): Hela (blue). Primary Antibody (green line): Rabbit Anti-Mtp53 (N235K N239Y) antibody (orb11211), 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 1 X 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.

Fixative-fixed, paraffin embedded (Human cervical carcinoma), 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 (Mtp53 (N235K N239Y)) Polyclonal Antibody, Unconjugated (orb11211) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 minutes and DAB staining.

Tissue/Cell: Hela, 4% Fixative-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Mtp53 (N235K N239Y)) Polyclonal Antibody, Unconjugated (orb11211) 1:200, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.