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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...

Proteomics

Proteomics

A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
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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.
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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: Jun 6, 2026

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Published on: April 2, 2018

Gel-based and gel-free proteomic technologies.

Peter Scherp¹, Ginger Ku, Liana Coleman

¹Proteomics and Metabolomics Core, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA.

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

|November 18, 2010

Summary

This study presents detailed protocols for proteomic analysis of adipose-derived stem cells (ASCs). It highlights mass spectrometry (MS)-based methods, including 2D-DIGE-MS and 2D-LC-MS, for protein identification and quantification.

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Last Updated: Jun 6, 2026

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

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Area of Science:

Proteomics and Mass Spectrometry
Stem Cell Biology
Biochemistry

Background:

Proteomics analyzes protein expression, localization, function, modifications, and interactions.
Mass spectrometry (MS)-based methods are crucial for high-throughput protein identification and quantification.
Understanding protein changes is key to cellular and physiological processes.

Purpose of the Study:

To provide detailed protocols for global proteomic analysis of adipose-derived stem cells (ASCs).
To present two central strategies: 2D-DIGE-MS and 2D-LC-MS.
To enable systematic and unbiased identification and quantification of complex protein mixtures.

Main Methods:

Utilized gel-based (2D-DIGE) and gel-free (2D-LC) separation techniques prior to MS analysis.
Applied mass spectrometry for comprehensive protein profiling.
Developed and combined various electrophoretic and chromatographic methods.

Main Results:

Established detailed protocols for proteomic analysis of ASCs.
Demonstrated the utility of 2D-DIGE-MS and 2D-LC-MS for ASC proteome.
Enabled the potential discovery of novel protein interactions and cellular changes.

Conclusions:

The presented protocols facilitate in-depth proteomic studies of ASCs.
MS-based proteomics offers powerful tools for discovering cellular regulatory mechanisms.
These methods are vital for advancing stem cell research and understanding physiological processes.