Horseradish peroxidase hrp conjugated anti mouse antibody

The expression of COL II in the engineered cartilage was examined by immunohistochemical staining at 12 and 24 weeks after implantation. Briefly, paraffin sections were deparaffinised followed by hydration in ethanol solutions of decreasing concentrations (100% to 70%), and pre-treated with 0.1% of Proteinase K (Vivantis Technologies, Oceanside, California) at room temperature for 30 minutes. The sections were then treated with protein blocking agent ( UltraTech HRP Kit, Beckman Coulter France, Villepinte, France) for ten minutes and incubated with goat serum to block non-specific sites. The slides were washed in PBS three times and the sections were incubated at 37°C for one hour with mouse anti-collagen type II monoclonal antibody diluted in PBS (1:200), followed by incubation with 1:100 diluted horseradish peroxidase (HRP)-conjugated anti-mouse antibody (Dako, Carpinteria, California) for 30 minutes. Colour was developed with diaminobenzidine tetrahydrochloride (DAB).