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Related Concept Videos

Two-dimensional Gel Electrophoresis01:22

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-PAGE01:27

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...
DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Capillary Electrophoresis: Applications01:30

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.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...

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Related Experiment Video

Updated: Jul 5, 2026

Proteomic Profiling of Macrophages by 2D Electrophoresis
07:53

Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

Protein profiling using two-dimensional difference gel electrophoresis (2-D DIGE).

Kathryn S Lilley1

  • 1University of Cambridge, Cambridge, United Kingdom.

Current Protocols in Protein Science
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

Two-dimensional difference gel electrophoresis (2D-DIGE) uses fluorescent dyes for precise protein quantification. This method simplifies comparative proteomics by enabling direct spot matching within a single gel, improving accuracy in identifying expression differences.

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Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting
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Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome
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Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

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Last Updated: Jul 5, 2026

Proteomic Profiling of Macrophages by 2D Electrophoresis
07:53

Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting
10:51

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

Published on: April 10, 2014

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome
12:34

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

Published on: April 2, 2018

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • Comparative proteomics requires accurate quantification of protein expression levels.
  • Traditional 2D gel electrophoresis faces challenges in inter-gel spot matching and quantification.

Purpose of the Study:

  • To describe the 2D Difference Gel Electrophoresis (2D-DIGE) procedure.
  • To detail sample preparation, protein labeling, and downstream processing for 2D-DIGE.

Main Methods:

  • Pre-electrophoretic labeling of protein samples with spectrally distinct fluorescent dyes.
  • Running all labeled samples in a single 2D gel.
  • Scanning the gel at different wavelengths to view each dye-labeled sample individually.
  • Utilizing image analysis software to quantify spot intensities and determine expression ratios.

Main Results:

  • 2D-DIGE circumvents spot matching issues inherent in traditional 2D gel electrophoresis.
  • The method allows for accurate identification and quantification of protein expression differences.
  • Successful application demonstrated across various cell types.

Conclusions:

  • 2D-DIGE is a robust technique for quantitative comparative proteomics.
  • The described protocol facilitates reliable protein expression analysis.
  • This method enhances the accuracy and efficiency of differential protein expression studies.