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Technical Aspects of Immunohistochemistry

Animal Disease Diagnostic Laboratory and Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN

Immunohistochemistry is an integral technique in many veterinary laboratories for diagnostic and research purposes. In the last decade, the ability to detect antigens (Ags) in tissue sections has improved dramatically, mainly by countering the deleterious effects of formaldehyde with antigen retrieval (AR) and increasing sensitivity of the detection systems. In this review, I address these topics and provide an overview of technical aspects of immunohistochemistry, including those related to antibodies (Abs) and Ags, fixation, AR, detection methods, background, and troubleshooting. Microarray technology and the use of rabbit monoclonal Abs in immunohistochemistry are also discussed.

Key words: Antibodies; antigen retrieval; antigens; detection methods; fixation; immunohistochemistry.

Request reprints from Dr. José Ramos-Vara, Animal Disease Diagnostic Laboratory, Purdue University, 406 South University, West Lafayette, IN 47907 (USA). E-mail: ramosja{at}purdue.edu

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