Related Experiment Video

Updated: May 15, 2026

Histological-Based Stainings Using Free-Floating Tissue Sections
06:45

Histological-Based Stainings Using Free-Floating Tissue Sections

Published on: August 25, 2020

Immunostaining: detection of signaling protein location in tissues, cells and subcellular compartments

Biswanath Maity1, David Sheff, Rory A Fisher

  • 1Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA, USA.

Methods in Cell Biology
|January 16, 2013
PubMed

Insights

Immunostaining uses antibodies to detect specific proteins in cells or tissues. This protocol details immunocytochemistry and immunohistochemistry methods, including immunofluorescence and chemiluminescence for visualization.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Immunology

Background:

  • Immunostaining is a crucial technique for visualizing specific proteins within biological samples.
  • Understanding protein distribution and localization is vital for deciphering cellular functions and disease mechanisms.

Purpose of the Study:

  • To detail methodologies for protein detection and localization in cells and tissues using immunostaining.
  • To describe the principles and applications of immunocytochemistry and immunohistochemistry.
  • To outline the use of immunofluorescence and chemiluminescence for visualizing antigen-antibody complexes.

Main Methods:

  • Utilizes specific primary antibodies to bind target proteins in fixed cells or tissues.
  • Employs secondary antibodies for detection, often conjugated to fluorophores or enzymes.
  • Details protocols for both immunocytochemistry (cells) and immunohistochemistry (tissues).

Main Results:

  • Provides a comprehensive guide to performing immunostaining techniques.
  • Explains the visualization of protein localization at cellular and subcellular levels.
  • Highlights the differential detection of proteins using immunofluorescence and chemiluminescence.

Conclusions:

  • Immunostaining is a versatile method for studying protein expression and localization.
  • The described protocols enable detailed analysis of protein distribution in various biological contexts.
  • Accurate protein localization through immunostaining aids in understanding cellular processes and disease pathology.

More Related Videos

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections09:11

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

Published on: November 14, 2016

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging08:40

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Published on: April 8, 2016

Related Concept Videos

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.
Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...
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...