Mouse monoclonal anti p53 antibody

According to our previous study (11 (link)), immunohistochemistry assay was done. Briefly, formalin-fixed sections were rehydrated using the graded serious ethanol solution. Antigen retrieval was performed in the 10 mM citrate buffer at 98–100 °C for 30 min and 10% normal goat serum was used to block the unspecific binding for 30 min. The primary antibodies including mouse monoclonal anti-GAGE7 antibody (1:100 dilution; Abcam, UK), mouse monoclonal anti-MAGEA1 antibody (1:100 dilution; Absin, China), mouse monoclonal anti-PGP9.5 antibody (1:200 dilution; Abcam, UK), mouse monoclonal anti-CAGE antibody (1:100 dilution; Absin, China) and mouse monoclonal anti-p53 antibody (1:200 dilution; Abcam, UK) were added on the section overnight at 4 °C, followed by incubation with HRP-conjugated goat anti-mouse secondary antibody for 30 min at room temperature (Sigma-Aldrich, Poole, Dorset, UK).
Immunohistochemical evaluation was graded based on the intensity of staining (1+, weak; 2+, moderate; 3+, strong) and the percentage of stained cells (0, for <5%; 1, for 5–25%; 2, for 26–50% and 3, >50%) (12 (link)). The score used for the analyses was the average values. When combining these two parameters, 0–1 and >1 were considered negative and positive staining, respectively. Scoring was reviewed in parallel by two experienced pathologists who were blinded to all clinical data.