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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.

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Related Experiment Video

Updated: Mar 14, 2026

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

A bead-based western for high-throughput cellular signal transduction analyses.

Fridolin Treindl^1,2, Benjamin Ruprecht^3,4, Yvonne Beiter^1,2

¹NMI Natural and Medical Sciences Institute at the University of Tübingen, 72770 Reutlingen, Germany.

Nature Communications

|September 24, 2016

Summary

The DigiWest approach enhances western blotting for proteomic analysis of limited cell samples. This high-throughput method identified key signaling changes during Lapatinib resistance, involving Ephrin and p53 pathways.

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Last Updated: Mar 14, 2026

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

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Published on: September 26, 2020

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

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Published on: February 5, 2014

A Western Blotting Protocol for Small Numbers of Hematopoietic Stem Cells

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Published on: August 22, 2018

Area of Science:

Proteomics
Cellular Signalling
Biotechnology

Background:

Analyzing large numbers of proteins is crucial for dissecting cellular signalling pathways.
Traditional western blotting has limitations in throughput and sample requirements for comprehensive analysis.

Purpose of the Study:

To introduce and validate the DigiWest approach, a bead-based microarray platform for high-throughput western blotting.
To enable comprehensive proteomic-scale analysis of limited biological samples.
To investigate alterations in cellular signal transduction during the development of resistance to Lapatinib.

Main Methods:

Developed the DigiWest approach by combining gel-based protein separation with immobilization on microspheres.
Created hundreds of blot replicas on a bead-based microarray platform.
Utilized an automated platform for rapid quantification of hundreds of proteins and their modifications in complex samples.

Main Results:

Demonstrated high-throughput western blotting with molecular weight resolution, sensitivity, and signal linearity.
Successfully analyzed limited material, such as cells from laser capture microdissection.
Identified significant alterations in cellular signal transduction during Lapatinib resistance, including Ephrin-mediated signalling and p53-controlled processes.

Conclusions:

The DigiWest approach extends traditional western blotting to proteomic scales, enabling comprehensive analysis of limited samples.
This high-throughput method facilitates rapid quantification of numerous proteins and modifications in complex biological systems.
Revealed critical changes in signalling pathways, highlighting the role of Ephrin and p53 in Lapatinib resistance.