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.

Southern Blot

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...

Two-dimensional Gel Electrophoresis

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such...

You might also read

Related Articles

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

Sort by

Same author

Multiple analyte profiling (MAP) index as a powerful diagnostic and therapeutic monitoring tool.

Methods (San Diego, Calif.)·2020

Same author

Method to quantify cytokines and chemokines in mouse brain tissue using Bio-Plex multiplex immunoassays.

Methods (San Diego, Calif.)·2019

Same author

Multiplexing techniques for measurement of the immunomodulatory effects of particulate materials: Precautions when testing micro- and nano-particles.

Methods (San Diego, Calif.)·2019

Same author

V3 stain-free workflow for a practical, convenient, and reliable total protein loading control in western blotting.

Journal of visualized experiments : JoVE·2014

Search research articles

Related Experiment Video

Updated: Nov 22, 2025

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

Published on: February 5, 2014

Technical Considerations for Contemporary Western Blot Techniques.

¹Bio-Rad Laboratories, Hercules, CA, USA. Kenneth_Oh@bio-rad.com.

Methods in Molecular Biology (Clifton, N.J.)

|January 9, 2021

Summary

Western blotting remains a vital laboratory technique. Modern advancements in antibodies, detection methods, and imaging enhance its sensitivity, dynamic range, and user-friendliness for improved experimental outcomes.

Keywords:

Binning Bit depth CCD Chemiluminescence Digital imaging Dynamic range Enhanced chemiluminescence Fluorescence Housekeeping normalization Immunoblotting Immunodetection Multiplex Quantitation Resolution Total protein normalization Western blot

More Related Videos

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

Related Experiment Videos

Last Updated: Nov 22, 2025

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

Published on: February 5, 2014

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

Area of Science:

Biochemistry
Molecular Biology
Immunology

Background:

Western blotting is a widely used laboratory technique for protein analysis.
Despite its age (over 40 years), its utility, low cost, and accessibility ensure its continued relevance.
Ongoing advancements are improving the technique's performance and ease of use.

Purpose of the Study:

To discuss contemporary advancements in western blotting.
To guide users in leveraging these improvements for western blot optimization.
To highlight strategies for maximizing sensitivity, dynamic range, and usability.

Main Methods:

Review of recent developments in antibody technology for enhanced specificity.
Discussion of updated chemiluminescent and fluorescent detection systems.
Analysis of modern imaging techniques for improved western blot analysis.

Main Results:

Newer antibodies offer increased specificity, reducing background noise.
Advanced detection reagents provide greater sensitivity and broader dynamic range.
Modern imaging systems facilitate easier data acquisition and analysis.

Conclusions:

Western blotting continues to evolve with technological progress.
Optimizing antibody selection, detection methods, and imaging is crucial for maximizing assay performance.
These contemporary practices enhance the reliability and efficiency of western blot experiments.