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Heat-Induced Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections

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Immunocytochemical Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 588))

Abstract

The development of heat-induced antigen (epitope) retrieval (HIER) technologies has led to dramatic improvements in our ability to detect antigens in formalin fixed, archival tissues. Paradoxically, wet heat treatment at temperatures greater than 95°C in appropriate buffer solutions can reconstitute the antigenicity of many proteins that have been rendered nonreactive during the fixation and paraffin embedding process, which heretofore could only be identified in fresh or frozen tissues. The reason for this effect is unclear, but it has been suggested that the vigorous heat treatment partially reverses or disrupts the aldehyde cross-links occurring in proteins during formalin fixation and restores the original conformation of antigenic epitopes. The great success of antigen/epitope retrieval technologies further emphasizes the importance of preanalytical steps in immunohistochemistry. Over the past several years, since this technology was first reported, there have been numerous modifications to the original formulation. It is the purpose of this chapter to discuss the critical issues required for optimal HIER and to provide guidelines for the use of popular HIER buffers and heating devices used for routine immunohistochemical detection.

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Authors and Affiliations

  1. Laboratory of Tumor Pathology and Molecular Diagnostics, Bay Zoltan Foundation for Applied Research, Szeged, Hungary

    Laszlo Krenacs & Eva Stelkovics

  2. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary

    Tibor Krenacs

  3. Laboratory of Pathology, Hematopathology Section, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Mark Raffeld

Authors
  1. Laszlo Krenacs
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  2. Tibor Krenacs
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  3. Eva Stelkovics
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  4. Mark Raffeld
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Corresponding author

Correspondence to Tibor Krenacs .

Editor information

Editors and Affiliations

  1. Fac. Medicina de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes 3900, Ribeirão Preto, 14049-900, Brazil

    Constance Oliver

  2. Fac. Medicina de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes 3900, Ribeirão Preto, 14049-900, Brazil

    Maria Célia Jamur

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Krenacs, L., Krenacs, T., Stelkovics, E., Raffeld, M. (2010). Heat-Induced Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections. In: Oliver, C., Jamur, M. (eds) Immunocytochemical Methods and Protocols. Methods in Molecular Biology, vol 588. Humana Press. https://doi.org/10.1007/978-1-59745-324-0_14

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  • DOI: https://doi.org/10.1007/978-1-59745-324-0_14

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-463-0

  • Online ISBN: 978-1-59745-324-0

  • eBook Packages: Springer Protocols

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