Immunohistochemistry

Immunohistochemistry (IHC) is the technique of assessing the expressed protein in vivo through antigen-antibody reaction. In this technique, a more expressed protein is darkly stained than a less expressed one. Immunohistochemistry (IHC) is a technique that involves histological, immunological, and biochemical techniques. IHC with both direct (involving primary antibody) and indirect (involving both primary and secondary antibodies) methods was discussed. A detailed protocol for IHC has been discussed with practical examples of IHC as conducted in our laboratory of growth hormone in pituitary gland, and CD4 and CD8 expressions in uterus. The basic steps for IHC include fixing and embedding the tissue; cutting and mounting the section; deparaffinizing and rehydrating the section; antigen retrieval; immunohistochemical staining; counterstaining (if desired); dehydrating and stabilizing with mounting medium; and viewing the staining under the microscope.