1. Prepared 18 micron frozen rat brain tissue sections (post fixed with 10% formalin for 30 minutes).
2. Dilute the primary antibody 1:100 and incubated with diluted FIH antibody (cat# NB 100-428) at 4C, overnight.
3. Diluted the secondary antibody, biotinylated anti-rabbit IgG, according to the manufacturer's guidelines and incubated overnight at 4C.
4. Diluted the tertiary antibody, HRP-conjugated streptavidin, according to the manufacturer's guidelines and incubated overnight at 4C.
5. Visualized with DAB oxidation by HRP.
