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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 as cells...

Electrophoresis: Overview

Electrophoresis: Overview

Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...

Capillary Electrophoresis: Instrumentation

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...

SDS-PAGE

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...

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Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
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Related Experiment Video

Updated: Jul 11, 2026

High-throughput Protein Expression Generator Using a Microfluidic Platform

High-throughput Protein Expression Generator Using a Microfluidic Platform

Published on: August 23, 2012

Microfabricated two-dimensional electrophoresis device for differential protein expression profiling.

Charles A Emrich¹, Igor L Medintz, Wai K Chu

¹Biophysics Graduate Group, University of California, Berkeley, California 94720, USA.

Analytical Chemistry

|September 8, 2007

Summary

A novel microfluidic system enables reproducible two-dimensional differential gel electrophoresis (DIGE) for analyzing complex protein mixtures from E. coli. This micro-DIGE analyzer advances protein analysis in microfabricated devices.

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Last Updated: Jul 11, 2026

High-throughput Protein Expression Generator Using a Microfluidic Platform

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Published on: August 23, 2012

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Published on: November 4, 2014

Digital Microfluidics for Automated Proteomic Processing

Digital Microfluidics for Automated Proteomic Processing

Published on: November 6, 2009

Area of Science:

Proteomics
Analytical Chemistry
Biotechnology

Background:

Two-dimensional gel electrophoresis (2D-PAGE) is a powerful technique for protein separation.
Microfluidic devices offer advantages in sample handling and analysis speed for proteomics.

Purpose of the Study:

To develop a microfluidic system for two-dimensional differential gel electrophoresis (DIGE).
To demonstrate reproducible separation of complex protein mixtures using the micro-DIGE analyzer.

Main Methods:

A two-layer borosilicate glass microdevice was fabricated for isoelectric focusing and native electrophoresis.
A microfluidic interface (MFI) connected the orthogonal separation channels.
Proteins were labeled with Cy2 and Cy3 dyes and detected via fluorescence scanning.
7 M urea was used in the second-dimension matrix for reproducibility.

Main Results:

The micro-DIGE analyzer successfully performed reproducible 2D separations of purified proteins and E. coli protein mixtures.
The system tracked induced expression of maltose binding protein.
Minimal run-to-run variation was achieved.

Conclusions:

The micro-DIGE analyzer represents a significant advancement for reproducible 2D protein analysis in microdevices.
The developed microchannel interface structures can be applied to other multidimensional microfluidic separations.