Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

6.0K
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...
6.0K
SDS-PAGE01:27

SDS-PAGE

23.2K
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...
23.2K
Electrophoresis: Overview01:20

Electrophoresis: Overview

4.1K
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...
4.1K
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

1.9K
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,...
1.9K
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

6.2K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
6.2K
DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

96.0K
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...
96.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The spatial proteome of the Plasmodium falciparum schizont illuminates the composition and evolutionary trajectories of its organelles.

Nature communications·2026
Same author

Dynamic subcellular proteomics identifies regulators of adipocyte insulin action.

Nature communications·2026
Same author

Subcellular localization as a driver of protein function.

Nature reviews. Molecular cell biology·2026
Same author

Functional characterisation of tumour suppressor PDCD4 reveals previously undisclosed role in the control of cell adhesion.

Nucleic acids research·2026
Same author

Semi-supervised Bayesian integration of multiple spatial proteomics datasets.

PLoS computational biology·2025
Same author

The distinct roles of genome, methylation, transcription, and translation on protein expression in Arabidopsis thaliana resolve the Central Dogma's information flow.

Genome biology·2025
Same journal

De Novo Protein Design Using the Blueprint Builder in Rosetta.

Current protocols in protein science·2020
Same journal

Methods for Expression of Recombinant Proteins Using a Pichia pastoris Cell-Free System.

Current protocols in protein science·2020
Same journal

Histone Purification Combined with High-Resolution Mass Spectrometry to Examine Histone Post-Translational Modifications and Histone Variants in Caenorhabditis elegans.

Current protocols in protein science·2020
Same journal

A Rapid and Facile Purification Method for Glycan-Binding Proteins and Glycoproteins.

Current protocols in protein science·2020
Same journal

Synthesis of Recombinant Human Hemoglobin With NH<sub>2</sub> -Terminal Acetylation in Escherichia coli.

Current protocols in protein science·2020
Same journal

A Fluorescence-Based Assay to Monitor SUMOylation in Real-Time.

Current protocols in protein science·2020
See all related articles

Related Experiment Video

Updated: May 3, 2026

Proteomic Profiling of Macrophages by 2D Electrophoresis
07:53

Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

12.5K

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

Renata Feret1, Kathryn S Lilley1

  • 1Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.

Current Protocols in Protein Science
|February 11, 2014
PubMed
Summary
This summary is machine-generated.

Two-dimensional Difference Gel Electrophoresis (2-D DIGE) uses fluorescent dyes for precise protein quantification. This method minimizes gel-to-gel variability, enabling accurate identification of protein abundance changes.

Keywords:
2-D gel electrophoresisDIGEproteomicsquantitative

More Related Videos

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

13.1K
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

15.5K

Related Experiment Videos

Last Updated: May 3, 2026

Proteomic Profiling of Macrophages by 2D Electrophoresis
07:53

Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

12.5K
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

13.1K
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

15.5K

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • Two-dimensional gel electrophoresis is a standard technique for protein separation.
  • Inter-gel variability and spot matching challenges can complicate quantitative analysis.

Purpose of the Study:

  • To describe the 2-D DIGE procedure for accurate protein quantification.
  • To detail sample preparation, fluorescent labeling, and downstream processing for 2-D DIGE.

Main Methods:

  • Pre-electrophoretic labeling of samples with spectrally distinct fluorescent dyes.
  • Running all labeled samples in a single 2-D gel.
  • Scanning the gel at different wavelengths to view individual dye-labeled samples.
  • Utilizing image analysis software to generate spot volume ratios for quantification.

Main Results:

  • 2-D DIGE circumvents gel-to-gel variation and spot matching issues.
  • Accurate identification and quantification of protein abundance level changes are enabled.
  • The method allows for comparative proteomic analysis across multiple samples.

Conclusions:

  • 2-D DIGE is a robust technique for quantitative proteomics.
  • The described protocol facilitates reliable protein abundance analysis from various cell types.
  • Careful downstream processing is crucial for accurate results in 2-D DIGE.