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Chapter 6 - Immunohistochemistry for Research Applications

Published online by Cambridge University Press:  16 June 2022

By
Trung Nguyen
Edited by
Trung Nguyen
Trung Nguyen
Affiliation:
Peter MacCallum Cancer Centre, Australia
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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.

Keywords

protocolantibody concentrateparametersready-to-use (RTU)dilutiontitreoptimizationspeciesepitope retrievaldetection
Type
Chapter
Information
Immunohistochemistry
A Technical Guide to Current Practices
 , pp. 160 - 182
Publisher: Cambridge University Press
Print publication year: 2022

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