Immunohistochemistry protocol specific for TRPV1 Antibody (NB300-122)

Procedure Guide for NB 300-122 Polyclonal Anti-Vanilloid Receptor I

Immunohistochemistry Procedure

1) Animals were perfused transcardially with calcium-free Tyrode's solution, followed by fixative, and finally with 10% sucrose in PBS. Antiserum was used on perfusion-fixed tissues.
2) Tissues were dissected and stored in PBS/10% sucrose, overnight.
3) Frozen tissues were sectioned and mounted on gelatin-coated slides.
4) Slide-mounted tissue was processed for indirect immunofluorescence. Then the slides were incubated with blocking buffer for 1 hour at room temperature.
5) NB300-122 (anti-Vanilloid Receptor I) was diluted in blocking buffer to 1:2000.
6) Blocking buffer was removed and diluted NB300-122 was added. The primary antibody incubation was done at 4 degrees C for 18-24 hours.
7) Slides were rinsed 3 times and then incubated with appropriate secondary antibody for 1 hour at room temperature.
8) Slides were rinsed 3 times.
9) Slides were coverslipped.
10) Staining was examined by fluorescent microscopy.