Abstract
Immunohistochemical techniques detect antigens in tissue sections by means of immunological and chemical reactions. This technique is highly sensitive and specific and can detect a wide variety of antigens in multiple animal species. This chapter reviews common immunohistochemical methods used in the characterization of normal and pathologic tissue and the reagents used. Pretreatments such as blocking steps for endogenous activities and antigen retrieval are included. Standard procedures on formalin-fixed, paraffin-embedded tissues as well as method standardization for new antibodies and troubleshooting are emphasized.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ramos-Vara, J.A., Kiupel, M., Baszler, T., Bliven, L., Brodersen, B., Chelack, B., Czub, S., Del Piero, F., Dial, S., Ehrhart, E.J., Graham, T., Manning, L., Paulsen, D., Valli, V.E., and West, K. (2008) Suggested guidelines for immunohistochemical techniques in veterinary diagnostic laboratories. J. Vet. Diagn. Invest. 20, 393–413.
Ramos-Vara, J.A., and Beissenherz, M.E. (2000) Optimization of immunohistochemical methods using two different antigen retrieval methods on formalin-fixed, paraffin-embedded tissues: experience with 63 markers. J. Vet. Diagn. Invest. 12, 307–311.
Ramos-Vara, J.A. (2005) Technical aspects of immunohistochemistry. Vet. Pathol. 42, 405–426.
Van der Loos, C.M. (1999) Immunoenzyme Multiple Staining Methods, Bios Scientific Publishers, Oxford, UK.
Atkins, D., Reiffen, K.-A., Tegtmeier, C.L., Winther, H., Bonato, M.S., and Störkel, S. (2004) Immunohistochemical detection of EGFR in paraffin-embedded tumor tissues: variation in staining intensity due to choice of fixative and storage time of tissue sections. J. Histochem. Cytochem. 52, 893–901.
Fergenbaum, J.H., Gracia-Closas, M., Hewitt, S.M., Lissowska, J., Sakoda, L.C., and Sherman, M.E. (2004) Loss of antigenicity in stored sections of breast cancer tissue microarrays. Cancer Epidemiol. Biomarkers Prev. 13, 667–672.
Mirlacher, M., Kasper, M., Storz, M., Knecht, Y., Durmuller, U., Simon, R., Mihatsch, M.J., and Sauter, G. (2004) Influence of slide aging on results of translational research studies using immunohistochemistry. Mod. Pathol. 17, 1414–1420.
Blind, C., Koepenik, A., Pacyna-Gengelbach, M., Fernahl, G., Deutschmann, N., Dietel, M., Krenn, V., and Petersen, I. (2008) Antigenicity testing by immunohistochemistry after tissue oxidation. J. Clin. Pathol. 61, 79–83.
Wood, G.S., and Warnke, R. (1981) Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems. J. Histochem. Cytochem. 29, 1196–1204.
Henwood, A.F. (2005) Effect of slide drying at 80°C on immunohistochemistry. J. Histotechnol. 28, 45–46.
Malik, N.J., and Daymon, M.E. (1982) Improved double immunoenzyme labeling using alkaline phosphatase and horseradish peroxidase. J. Clin. Pathol. 35, 1092–1094.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ramos-Vara, J.A. (2011). Principles and Methods of Immunohistochemistry. In: Gautier, JC. (eds) Drug Safety Evaluation. Methods in Molecular Biology, vol 691. Humana Press. https://doi.org/10.1007/978-1-60761-849-2_5
Download citation
DOI: https://doi.org/10.1007/978-1-60761-849-2_5
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60327-186-8
Online ISBN: 978-1-60761-849-2
eBook Packages: Springer Protocols