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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...

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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Updated: May 27, 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

2-D gel electrophoresis: constructing 2D-gel proteome reference maps.

Maria Paola Simula¹, Agata Notarpietro, Giuseppe Toffoli

¹Experimental and Clinical Pharmacology Unit, CRO Centro di Riferimento Oncologico, IRCCS National Cancer Institute, via F. Gallini 2, Aviano-PN 33081, Italy. mariapaola.simula@aots.sanita.fvg.it

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

|December 2, 2011

Summary

This study presents the first two-dimensional gel electrophoresis (2-DE) reference map of the human small intestine. This advanced proteomic map, generated using 2D-Difference in Gel Electrophoresis (2D-DIGE), aids in understanding cellular protein expression and disease mechanisms.

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Area of Science:

Proteomics
Biochemistry
Molecular Biology

Background:

Two-dimensional gel electrophoresis (2-DE) is a key technique for protein separation based on isoelectric point (pI) and molecular mass.
2D-Difference in Gel Electrophoresis (2D-DIGE) enhances conventional 2-DE for more accurate quantitative proteomic analysis.
Detailed 2D reference maps are valuable for studying protein expression changes in biological systems and disease.

Purpose of the Study:

To establish the first comprehensive 2-DE reference map of the human small intestine.
To provide a foundational resource for proteomic studies of the human small intestine.
To facilitate the identification of molecular bases of pathological processes in the small intestine.

Main Methods:

Utilized 2D-Difference in Gel Electrophoresis (2D-DIGE) technology for high-resolution protein separation.
Applied established 2-DE protocols to human small intestine tissue samples.
Generated a detailed protein map based on isoelectric focusing and SDS-PAGE.

Main Results:

Successfully created the first 2-DE reference map of the human small intestine proteome.
The map provides a detailed overview of protein distribution based on pI and molecular mass.
Demonstrated the utility of 2D-DIGE for generating comprehensive proteomic reference maps.

Conclusions:

The developed 2-DE reference map serves as a crucial resource for human small intestine research.
This map can aid in understanding normal protein homeostasis and identifying disease-related alterations.
The protocol provides a robust method for generating reference maps in other biological systems.