Immunohistochemistry: techniques and considerations. Nina Cluny January 2010. Immunohistochemistry. What is it? What can it tell you? What are its limitations? How is it done? What are the controls? Quantification Single vs double or triple labelling. Paratope + Epitope. Ab-Ag complex.
This distinction is often overlooked.
Prevention of Ag translocation and/or degradation
1. Preabsorb the primary antibody with the immunizing antigen (question a company which does not supply this)
2. Replace the primary with preimmune serum from the same animal that the primary was raised in – or control ascites fluid for monoclonals
PPARa immunoreactivity in rat ileum
Intrinsic primary afferent neurons of the myenteric plexus are Dogiel Type II neurons that express calbindin
Red (CY3): rat anti-substance P
Green (FITC): mouse anti-VIP
Blue (AMCA): rabbit anti-CGRP
Blue - DAPI; Green (FITC): rabbit anti-CART; Red (CY3): goat anti-leptin; Purple (false
coloured, CY5): rat anti-NPY