Goat anti myelin basic protein

On P6 and P30, rat pups were anesthetized and perfused transcardially with saline followed by 4% paraformaldehyde. The coronal brain blocks including the corpus callosum were dissected, and free-floating sections (20 µm) of the brain were sliced in 4% paraformaldehyde at 4 °C. The free-floating sections were incubated in 3% H₂O₂ to suppress endogenous peroxidase activity. After blocking in PBS containing 5% bovine serum albumin (BSA) and 0.4% Triton X-100 for 30 min at 37 °C, the sections were incubated with mouse anti-O4 (1:100, Sigma-Aldrich, St. Louis, MO, USA) antibody and goat anti-myelin basic protein (MBP) (1:200, Santa Cruz Biotechnology, San Francisco, CA, USA) overnight at 4 °C. The sections were then washed again with PBS and incubated with appropriate horseradish peroxidase-conjugated secondary antibodies (1:200, Santa Cruz) at 37 °C for 3 h. Finally, the immunoreaction products of the sections were visualized with a DAB kit (Zhongshan, Beijing, China) [23 (link)]. Negative controls were set up by omitting the primary antibodies.