Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Western Blotting

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

Immunogold Electron Microscopy

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.

Immunoprecipitation

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...

Immunocytochemistry and Immunohistochemistry

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...

Enzyme-Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.

Immunofluorescence Microscopy

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

First-in-Human Study of a Novel Coronary Crossing Device for Calcified Lesions.

JACC. Cardiovascular interventions·2025

Same author

Analysis of seat belt buckle release forces in post-rollover scenarios: Implications for passenger safety.

Applied ergonomics·2025

Same author

A fatigue failure framework for the assessment of highly variable low back loading using inertial motion capture - a case study.

Ergonomics·2025

Same author

Critical review of chronic toxicity testing approaches with the saltwater mysid (Americamysis bahia) used in pesticide registration.

Environmental toxicology and chemistry·2025

Same author

Scan/rescan reliability of magnetic resonance imaging (MRI).

European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society·2025

Same author

Expedited conveyance of out-of-hospital-cardiac arrest patients with STEMI and shockable rhythms to Cardiac Arrest Centres - A feasibility pilot study of the British Cardiovascular Intervention Society conveyance algorithm.

Resuscitation·2025

Same journal

A Comprehensive Experimental Guide to Studying Cross-Presentation in Dendritic Cells In Vitro.

Current protocols in immunology·2020

Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020

Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020

Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020

Same journal

Monoclonal Antibodies to Human Cell Surface Antigens.

Current protocols in immunology·2020

Same journal

Protocols for Experimental Sjögren's Syndrome.

Current protocols in immunology·2020

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 27, 2026

Immunoblot Analysis

Immunoblot Analysis

Published on: June 20, 2008

Immunoblotting and immunodetection.

Sean Gallagher¹, Scott E Winston Tank Transfer Systems², Steven A Fuller Tank Transfer Systems²

¹UVP, Inc, Upland, California.

Current Protocols in Immunology

|November 20, 2008

Summary

This summary is machine-generated.

Western blotting is a key technique for identifying specific antigens using antibodies. This protocol details protein solubilization, separation, transfer to membranes, and immunodetection for reliable results.

More Related Videos

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

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

Published on: November 14, 2016

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

Published on: January 5, 2024

Related Experiment Videos

Last Updated: Jun 27, 2026

Immunoblot Analysis

Immunoblot Analysis

Published on: June 20, 2008

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

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

Published on: November 14, 2016

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

Published on: January 5, 2024

Area of Science:

Biochemistry
Molecular Biology
Immunology

Background:

Immunoblotting, commonly known as western blotting, is a widely used technique.
It enables the identification of specific protein targets through antibody recognition.

Purpose of the Study:

To provide comprehensive protocols for western blotting.
To cover all essential steps from sample preparation to detection.

Main Methods:

Protein sample solubilization using SDS and reducing agents.
Separation of proteins via SDS-PAGE.
Electrophoretic transfer of proteins to a membrane.
Immunodetection using primary and secondary antibodies.
Detection via chromogenic or luminescent substrates.

Main Results:

Successful identification of specific antigens.
Demonstration of membrane stripping and reprobing capability.

Conclusions:

Western blotting is a versatile method for protein analysis.
The provided protocols ensure efficient and reliable antigen detection.