Related Experiment Video

Updated: Mar 8, 2026

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections
09:11

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

Published on: November 14, 2016

17.9K

Heat-Induced Antigen Retrieval in Immunohistochemistry: Mechanisms and Applications

Shuji Yamashita1, Osamu Katsumata2

  • 1Department of Pathology, School of Medicine, Keio University, 35-Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan. shuji@z5.keio.jp.

Methods in Molecular Biology (Clifton, N.J.)
|February 4, 2017
PubMed

Insights

Heat-induced antigen retrieval (HIAR) optimizes immunohistochemistry by detailing mechanisms and protocols for common tissue types. This essential technique enhances antigen visibility for accurate microscopic analysis.

Area of Science:

  • Biomedical Sciences
  • Histology
  • Immunohistochemistry

Background:

  • Heat-induced antigen retrieval (HIAR) is a critical step in immunohistochemistry.
  • It is essential for unmasking epitopes in fixed tissue specimens.
  • Standard protocols exist for various tissue preparations.

Purpose of the Study:

  • To elucidate the mechanisms underlying HIAR.
  • To provide comprehensive protocols for formalin-fixed, paraffin-embedded (FFPE) and frozen sections.
  • To describe advanced methods for challenging epitopes and multiplexed detection.

Main Methods:

  • Detailed description of HIAR mechanisms.
  • Protocols for FFPE and frozen tissue sections.
  • Methods for highly masked epitopes and double immunostaining (HRP/IF).

Main Results:

  • Established protocols for effective antigen retrieval in FFPE and frozen tissues.
  • Demonstrated successful retrieval of masked epitopes.
  • Validated double immunostaining techniques using HRP and immunofluorescence.

Conclusions:

  • HIAR is indispensable for modern immunohistochemistry.
  • The described protocols offer a robust framework for diverse applications.
  • Advanced methods enable sensitive detection of multiple antigens in single samples.

Keywords:
Double immunostainingFormalin-fixed, paraffin-embedded tissuesFrozen tissuesHRP-labeled antibody methodHeat-induced antigen retrievalHighly masked epitopesImmunofluorescence method

More Related Videos

Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections07:36

Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections

Published on: June 25, 2021

8.3K
Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment11:00

Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment

Published on: March 25, 2020

17.9K

Related Concept Videos

Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

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...
14.5K
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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.
5.7K