Monoclonal mouse anti ki67 antibody

Xenograft tissues were fixed with 4% paraformaldehyde (pH 7.4). For Ki67 staining, a monoclonal mouse anti-Ki67 antibody (Cell Signaling Technologies; 1:1000 dilution), a biotinylated secondary antibody (Sigma-Aldrich; 1:100 dilution), and DAB substrate system were used. Then sections were counter stained by hematoxylin. Images were analyzed with a NIKON ECLIPSE 80i microscope.

Standard procedure of Immunohistochemistry (IHC) analysis was performed. In brief, tissue sections (4 μm) were firstly deparaffinized, followed by citrate-based antigen retrieval and blocking with 0.2% endogenous peroxidase. After blocking with 10% bovine serum albumin (BSA, Sangon, Shanghai, China) for 45 min at room temperature, the slides were incubated with primary antibodies at 4 °C overnight. For Ki67 staining, sections from xenograft tumor tissues were incubated with a monoclonal mouse anti-Ki67 antibody (Cell Signaling Technology; 1:8,000 dilution; #2586), followed by staining with an HRP-conjugated secondary antibody (Abcam; 1:300 dilution; ab6728). Immunoreactivity was generated by the DAB system. Finally, sections were counterstained by hematoxylin, and images were analyzed with a NIKON microscope.