Principles of Effective Method Development in Pathobiology Research

Immunohistochemistry (IHC) is used to determine spatial relationships where we can identify the localization of target proteins in specific regions and cells of a tissue.  IHC has been the gold standard to analyze protein expression analysis in tissues; however, the IHC assay may not always work as not all antibodies are robust reagents for detection of target proteins.  For that reason, we have utilized several orthogonal assays to analyze localization of proteins and mRNA of targets within cells where robust antibody reagents are not available or specificity is in question. I will briefly describe several effective techniques that have helped my pathobiology team in the method development of assays that are used for target validation and characterization of human diseases and animal disease models.

Learning Objectives:

1. The presentation will provide the audience the many tools available for the method development of antibodies by utilizing orthogonal methods to ensure specificity of the IHC assay or equivalent histology-related assay

2. The presentation will help you discern how to resolve common problems encountered in immunohistochemical staining