Immunohistochemical Approaches for the Study of Spermatogenesis
Immunohistochemistry is an important technique that uses specific antibodies to determine the cellular localization of proteins/antigens in highly complex organs and tissues. While most immunohistochemistry experiments target protein epitopes, nonprotein antigens including BrdU may also be detected. Briefly, tissues are fixed, processed, sectioned, and then probed by a primary antibody while preserving the integrity of the tissue and cellular morphology. There are various methods available for visualization of the bound primary antibody that involve a reporter molecule which can be detected using light or fluorescent microscopy. Here we describe a basic immunohistochemistry protocol for identifying protein localization in testis sections using protein-specific antibodies.
Key wordsImmunohistochemistry Testis Antigen retrieval Antibody Antigen Biotin Fluorophores
The authors would like to thank Chris Small and Ryan Evanoff for their critical reading of the manuscript and Ryan for supplying the STRA8 immmunohistochemistry pictures. The work was supported by NIH grant HD RO1 10808 to MDG.
- 1.Coons A, Creech H, Jones R (1941) Immunological properties of an antibody containing a fluorescent group. Proc Soc Exp Biol Med 47:200–202Google Scholar
- 6.Russell LD et al (1990) Histological and histopathological evaluation of the testis, 1st edn. Cache River Press, St. Louis, MOGoogle Scholar