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Related Concept Videos

Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

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Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
These...
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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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Related Experiment Video

Updated: Feb 25, 2026

Primer for Immunohistochemistry on Cryosectioned Rat Brain Tissue: Example Staining for Microglia and Neurons
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Principles and Methods of Immunohistochemistry.

José A Ramos-Vara1

  • 1Department of Comparative Pathobiology, Purdue University, 406 South University, West Lafayette, IN, 47907, USA. ramosja@purdue.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 28, 2017
PubMed
Summary

Immunohistochemical techniques use immunological and chemical reactions to detect antigens in tissues. This review covers common methods, reagents, and standardization for characterizing normal and pathologic tissues.

Keywords:
Antigen retrievalDetection methodsFixationImmunohistochemistryStandardizationTroubleshooting

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Area of Science:

  • Histopathology
  • Immunology
  • Biochemistry

Background:

  • Immunohistochemistry (IHC) is a powerful tool for visualizing specific antigens within tissue samples.
  • It relies on the specific binding of antibodies to target antigens, followed by a detection system.

Purpose of the Study:

  • To provide a comprehensive review of common immunohistochemical methods.
  • To detail reagents and essential pretreatments for antigen detection.
  • To emphasize standardization and troubleshooting for reliable results.

Main Methods:

  • Review of established immunohistochemical protocols.
  • Discussion of reagent selection and optimization.
  • Inclusion of pretreatment steps like blocking and antigen retrieval.
  • Focus on formalin-fixed, paraffin-embedded tissues.

Main Results:

  • Detailed overview of sensitive and specific antigen detection across species.
  • Guidance on characterizing both normal and pathological tissue types.
  • Emphasis on reproducible results through method standardization.

Conclusions:

  • Immunohistochemistry is a versatile technique for diverse antigen detection in various species.
  • Proper method standardization and troubleshooting are crucial for accurate tissue characterization.
  • This review serves as a guide for implementing and optimizing IHC protocols.