Protocol specific for Neurokinin B Antibody (NB300-201)

Immunohistochemistry

1. Rats were perfused with 4% paraformaldehyde in 0.1M PBS.

2. The brains were then post-fixed in 4% paraformaldehyde and sucrose protected.

3. The treated sections were blocked in 0.1M Tris buffer containing 0.1% Triton X-100 and 0.25% BSA for 1 hour.

4. Anti-NKB (NB 300-201) was diluted in the blocking buffer and incubated with the sections overnight.

5. A goat anti-rabbit IgG conjugated to Alexa 488 was diluted in the blocking buffer and incubated with the sections overnight.

NOTE: Sections were made on a sledge microtome and stored in cryoprotectant solution at -20 degrees Celsius until ready to use.