Abstract
Immunohistochemistry (IHC) is a highly sensitive protein detection technique developed using the principle of antigen-antibody binding reaction. With immunohistochemistry, it is possible to visualize the abundance, distribution, and localization of proteins in situ. This chapter discusses the standard protocols involved in IHC detection using chromogenic substrates, including pre-treatment of tissues, types of chromogenic substrates, and troubleshooting at various stages of the protocol.
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References
Nakane PK, Pierce GB Jr (1966) Enzyme-labeled antibodies: preparation and application for the localization of antigens. J Histochem Cytochem 14:929–931
Mason DY, Sammons R (1978) Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. J Clin Pathol 31:454–460
Faulk WP, Taylor GM (1971) An immunocolloid method for the electron microscope. Immunochemistry 8:1081–1083
Gedda L, Bjorkelund H, Andersson K (2010) Real-time immunohistochemistry analysis of embedded tissue. Appl Radiat Isot 68:2372–2376
Werner M, Chott A, Fabiano A, Battifora H (2000) Effect of formalin tissue fixation and processing on immunohistochemistry. Am J Surg Pathol 24:1016–1019
Hewitson TD, Wigg B, Becker GJ (2010) Tissue preparation for histochemistry: fixation, embedding, and antigen retrieval for light microscopy. Methods Mol Biol 611:3–18
MacIntyre N (2001) Unmasking antigens for immunohistochemistry. Br J Biomed Sci 58:190–196
Jamur MC, Oliver C (2010) Permeabilization of cell membranes. Methods Mol Biol 588:63–66
Miller RT (2019) Avoiding pitfalls in diagnostic immunohistochemistry-important technical aspects that every pathologist should know. Semin Diagn Pathol 36:312–335
Ramos-Vara JA (2017) Principles and methods of immunohistochemistry. Methods Mol Biol 1641:115–128
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Vijayaraj, P. (2022). Immunochemistry Analysis Using Chromogenic Substrates on Tissue Sections. In: Ooi, A.T. (eds) Single-Cell Protein Analysis. Methods in Molecular Biology, vol 2386. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1771-7_1
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DOI: https://doi.org/10.1007/978-1-0716-1771-7_1
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1770-0
Online ISBN: 978-1-0716-1771-7
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