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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...

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Related Experiment Video

Updated: Jun 5, 2026

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

Published on: December 30, 2013

Coomassie staining as loading control in Western blot analysis.

Charlotte Welinder¹, Lars Ekblad

¹Department of Oncology, Clinical Sciences, Lund University, Sweden.

Journal of Proteome Research

|December 29, 2010

Summary

A new, faster protein staining method simplifies Western blotting analysis. This technique is compatible with standard blocking and can be used after immunodetection for improved results and further identification.

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Last Updated: Jun 5, 2026

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

Published on: December 30, 2013

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

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Published on: August 14, 2009

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Area of Science:

Biochemistry
Molecular Biology
Proteomics

Background:

Western blotting commonly uses housekeeping proteins to normalize electrophoresis loading.
Existing protein staining methods can be time-consuming and may interfere with subsequent analyses.

Purpose of the Study:

To develop a faster and simpler protein staining method for Western blotting.
To ensure compatibility with standard blocking conditions and immunodetection protocols.
To offer a method suitable for post-immunodetection analysis with enhanced linearity.

Main Methods:

A novel protein staining protocol was developed and tested.
Compatibility with standard blocking conditions was evaluated.
The method's performance after immunodetection was assessed for linearity.

Main Results:

The new staining method is significantly faster and simpler than conventional techniques.
It is fully compatible with ordinary blocking conditions used in Western blotting.
Post-immunodetection staining demonstrated superior linearity compared to existing methods.

Conclusions:

This improved protein staining technique offers a more efficient workflow for Western blotting.
Its compatibility and superior linearity make it a valuable tool for protein analysis.
The method allows for further identification of protein bands using peptide mass fingerprinting.