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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.

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...

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Updated: Jun 7, 2026

Digital Microfluidics for Automated Proteomic Processing

Digital Microfluidics for Automated Proteomic Processing

Published on: November 6, 2009

Automated microfluidic protein immunoblotting.

Mei He¹, Amy E Herr

¹Department of Bioengineering, University of California, Berkeley, California, USA.

Nature Protocols

|October 30, 2010

Summary

This study presents a rapid, automated method for native western blotting using photopatterned gels in microfluidic devices. This novel technique significantly reduces assay time and reagent consumption compared to traditional immunoblotting methods.

Area of Science:

Biochemistry
Analytical Chemistry
Microfluidics

Background:

Conventional immunoblotting methods are time-consuming, labor-intensive, and require significant reagent volumes.
Existing techniques lack seamless integration of multiple assay steps, hindering high-throughput analysis.
Limitations in conventional immunoblotting impede rapid protein analysis and biomarker discovery.

Purpose of the Study:

To develop a rapid and automated protocol for native western blotting using regional photopatterning of polyacrylamide gels.
To demonstrate the integration of multiple assay steps within a single microfluidic device.
To overcome the limitations of conventional immunoblotting, including manual intervention, low throughput, and high reagent consumption.

Main Methods:

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Digital Microfluidics for Automated Proteomic Processing

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Published on: November 6, 2009

Immunoblot Analysis

Immunoblot Analysis

Published on: June 20, 2008

Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood

Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood

Published on: April 16, 2016

Regional photopatterning of polyacrylamide gels within glass microfluidic devices.

Automated protein immunoblotting, specifically native western blotting, controlled by a high-voltage power supply.

Real-time monitoring of assay stages using an epifluorescence microscope and a charge-coupled device camera.

Main Results:

Successful implementation of rapid, automated native western blotting in microfluidic devices.
Fabrication and assay completion in approximately 3 hours, with the assay itself taking less than 5 minutes.
Significant reduction in manual steps, assay time, and reagent consumption compared to slab-gel immunoblotting.

Conclusions:

The described photopatterning technique provides a versatile platform for integrated bioanalytical assays.
This microfluidic approach offers substantial time savings and improved efficiency for protein immunoblotting.
The protocol forms a basis for developing advanced bioanalytical tools, including DNA/RNA blotting and multidimensional separations.