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Immunohistochemistry

Patricia A Loughran1,2, Mark A Ross1, Claudette M St Croix1

  • 1University of Pittsburgh, Center for Biologic Imaging, Pittsburgh, Pennsylvania.

Current Protocols
|September 14, 2022
PubMed

Insights

This guide details immunohistochemistry methods for protein localization in cells and tissues. It covers techniques for adherent and suspension cells, tissue sections, and troubleshooting antibody detection.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Histology

Background:

  • Immunohistochemistry is crucial for protein analysis in biological samples.
  • Accurate protein localization and quantification are vital in research and diagnostics.

Purpose of the Study:

  • To provide comprehensive protocols for protein labeling using immunohistochemistry.
  • To guide researchers in selecting antibodies and detection methods.
  • To offer troubleshooting advice for common issues in immunohistochemistry.

Main Methods:

  • Detailed protocols for immunofluorescent labeling of adherent and suspension cells.
  • Methods for immunofluorescent labeling of tissue sections.
  • Techniques for double-labeling using various antibody combinations and streptavidin-biotin conjugates.

Main Results:

  • Established protocols enable effective protein localization in diverse biological samples.
  • Guidance on antibody selection and detection systems optimizes experimental outcomes.
  • Troubleshooting tips address common challenges, improving reproducibility.

Conclusions:

  • This article serves as a practical resource for performing immunohistochemistry.
  • The provided methods facilitate accurate protein analysis in cellular and tissue contexts.
  • Effective application of these techniques supports advancements in biological research.

Keywords:
antibodies and fluorophoreimagingimmunofluorescencemicroscopy

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