Jove
Visualize
Contact Us

Related Concept Videos

Proteomics01:33

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.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
Diversity in Cell Signaling Responses01:22

Diversity in Cell Signaling Responses

The physiological function of a cell and cellular communication are outcomes of a range of extrinsic signals, intracellular signaling pathways, and cellular responses. No two cell types express the same repertoire of signaling components. Receptors are highly selective for their cognate ligands, but once activated, they can alter multiple cellular processes such as DNA transcription, protein synthesis, and metabolic activity. 
Graded and Abrupt Responses
Some signaling systems generate...
Protein Networks02:26

Protein Networks

An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
Interactions Between Signaling Pathways01:19

Interactions Between Signaling Pathways

Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...

You might also read

Related Articles

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

Sort by
Same author

Genetics of Childhood-Onset Systemic Lupus Erythematosus.

Arthritis & rheumatology (Hoboken, N.J.)·2025
Same author

Alterations in genes associated with cytosolic RNA sensing in whole blood are associated with coronary microvascular disease in SLE.

Scientific reports·2025
Same author

Alterations in genes associated with cytosolic RNA sensing in whole blood are associated with coronary microvascular disease in SLE.

Research square·2024
Same author

Reduced Left Ventricular Function on Cardiac MRI in SLE Patients Correlates with Measures of SLE Disease Activity and Inflammation.

Journal of radiology and clinical imaging·2024
Same author

Distinct RBC alloantibody responses in type 1 interferon-dependent and -independent lupus mouse models.

Frontiers in immunology·2024
Same author

Interaction effects of significant risk factors on low bone mineral density in ankylosing spondylitis.

PeerJ·2023
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 Experiment Video

Updated: Jun 23, 2026

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics
07:28

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics

Published on: October 19, 2021

2D-DIGE: comparative proteomics of cellular signalling pathways.

Nadia Ben Larbi1, Caroline Jefferies

  • 1RCSI Research Institute, Royal College of Surgeons in Ireland, Dublin 2, Ireland. nadiabenlarbi@rcsi.ie

Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
PubMed
Summary

Differential in-gel electrophoresis (DIGE) enables sensitive, comparative proteomic analysis by labeling samples with fluorescent dyes before running them on a single 2-D gel. This powerful technique aids in identifying protein expression changes and biological markers in various conditions.

More Related Videos

Navigating the Mass Spectrometry-Based Proteomic Data Using Free Computational Tools
07:01

Navigating the Mass Spectrometry-Based Proteomic Data Using Free Computational Tools

Published on: August 19, 2025

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

Related Experiment Videos

Last Updated: Jun 23, 2026

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics
07:28

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics

Published on: October 19, 2021

Navigating the Mass Spectrometry-Based Proteomic Data Using Free Computational Tools
07:01

Navigating the Mass Spectrometry-Based Proteomic Data Using Free Computational Tools

Published on: August 19, 2025

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

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • Two-dimensional (2-D) gel electrophoresis coupled with mass spectrometry (MS) is vital for comparative expression profiling of complex protein samples.
  • 2-D electrophoresis separates proteins by isoelectric point (pI) and molecular mass (Mr), offering a comprehensive proteome view.
  • Variations in protein expression, isoforms, and post-translational modifications can be identified, aiding in the discovery of biological markers for physiological or pathological states.

Purpose of the Study:

  • To describe the methods for differential in-gel electrophoresis (DIGE) for comparative proteomics.
  • To highlight DIGE's advantages in minimizing experimental variation and facilitating spot matching through simultaneous analysis of labeled samples on a single 2-D gel.
  • To detail protein labeling strategies, optimization of 2-D gel electrophoresis parameters, and protein preparation for MS identification.

Main Methods:

  • Differential in-gel electrophoresis (DIGE) using fluorescent dyes (e.g., Cy2, Cy3, Cy5) for labeling protein samples.
  • Separation of labeled proteins on a 2-D gel based on isoelectric point (pI) and molecular weight (MW).
  • Image capture and superposition of multiple fluorescent channels, followed by image analysis to identify differentially expressed protein spots.
  • Protein preparation from identified spots for subsequent mass spectrometry (MS) identification.

Main Results:

  • DIGE allows sensitive detection of proteins (down to 125 pg), with fluorescent dyes providing a linear response to protein concentration over five orders of magnitude.
  • Simultaneous analysis on a single gel minimizes variability and simplifies the matching of protein spots across different samples.
  • Identification of differentially regulated proteins, enabling comparative analysis of protein expression under various conditions (e.g., drug treatment, disease states).

Conclusions:

  • DIGE is a powerful tool for high-throughput comparative proteomics, offering enhanced sensitivity and reduced experimental variation.
  • The technique facilitates the identification of biological markers and the analysis of protein expression changes in response to stimuli or disease.
  • Detailed methods for DIGE, including labeling, electrophoresis, and MS preparation, are crucial for successful implementation.