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epitope retrieval

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Chapter 6 - Immunohistochemistry for Research Applications
- By Trung Nguyen
Edited by Trung Nguyen
Book:

Immunohistochemistry

Published online:

16 June 2022

Print publication:

07 July 2022, pp 160-182
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Summary

This chapter is written for the researcher who may encounter immunohistochemistry (IHC) in a slightly different context when compared to diagnostic applications. There are many moving parts to IHC assays, and this chapter covers all of the important aspects the researcher needs to consider when employing IHC for their projects. This objective is achieved by employing a request form for IHC services. The questions posed on the form build towards piecing together a protocol that is fit for purpose and can be used in many applications. Practical explanations about epitope retrieval, diluting antibodies from concentrates and the use of detection kits are provided. The need to block endogenous enzyme activity is also explained, as is the technique for antibody optimization. Borrowing the basic fundamental IHC protocol used in diagnostic histopathology, the researcher should be able to adopt and change parameters to suit their research applications.