IMMUNOCYTOCHEMISTRY FOR PARAFFIN-EMBEDDED SECTIONS

All steps done at room temp; can be done at 4°C if necessary but must be left overnight (for primary antibody step, e.g)

1. Wash sections 2x with 1xTBS, 10 minutes each wash
2. Add enough 0.1% Triton X in TBS to cover section, leave 10 minutes
3. Wash 2x with 1x TBS, 10 minutes each
4. Block with 5% BSA in TBS for 60 minutes
5. Add primary antibody in TBS + 0.8% BSA for 120 minutes
6. Wash 4x with TBS, 10 minutes each
7. Block for second antibody in normal serum (5% in TBS) of the host animal 15 minutes (e.g., if secondary antibody is made in goat, use goat serum; if donkey, use donkey serum, etc.)
8. Add secondary antibody (labeled e.g. with rhodamine) for 60 minutes
9. Wash 4 x in 1x TBS 10 minutes each
10. Cover slip with medium for non-dehydrated sections.