Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

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

Sort by

Same author

Addressing hurdles in cultured meat by exploring reduced myogenesis after bovine myoblast expansion.

Communications biology·2025

Same author

Quorum sensing in Saccharomyces cerevisiae brewing strains: effects of 2-phenylethanol on proteomic, lipidomic, and metabolomic profile.

FEMS yeast research·2025

Same author

Reliability of the International Spinal Cord Injury Physical Therapy-Occupational Therapy Basic Data Set.

Neurotrauma reports·2024

Same author

Chemo-Phosphoproteomic Profiling with ATR Inhibitors Berzosertib and Gartisertib Uncovers New Biomarkers and DNA Damage Response Regulators.

Molecular & cellular proteomics : MCP·2024

Same author

In vivo stress reporters as early biomarkers of the cellular changes associated with progeria.

Journal of cellular and molecular medicine·2022

Same author

CDKL5 kinase controls transcription-coupled responses to DNA damage.

The EMBO journal·2021

Search research articles

Related Experiment Video

Updated: Jun 22, 2026

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Published on: October 15, 2013

Protein detection and quantitation technologies for gel-based proteome analysis.

Walter Weiss¹, Florian Weiland, Angelika Görg

¹Technische Universität München, Fachgebiet Proteomik, Am Forum 2, D-85350 Freising-Weihenstephan, Germany.

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

|June 23, 2009

Summary

This review covers protein detection and quantitation in gel-based proteomics, focusing on universal, specific, and differential display methods. Fluorescent dye techniques are highlighted for their balance of sensitivity, quantitation, and practicality in proteomic analysis.

More Related Videos

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Published on: September 19, 2018

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

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 22, 2026

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Published on: October 15, 2013

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Published on: September 19, 2018

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

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
Analytical Chemistry

Background:

Gel-based proteomics relies on diverse protein detection and quantitation methods.
These methods fall into universal, specific post-translational modification detection, and differential display categories.
Key properties for proteomic analysis include sensitivity, dynamic range, reproducibility, cost, ease of use, and MS compatibility.

Purpose of the Study:

To review and categorize existing protein detection and quantitation methods in gel-based proteomics.
To highlight the advantages and limitations of various techniques, including staining and labeling.
To focus on fluorescent dye-based methods as a favored approach in current proteomic applications.

Main Methods:

Categorization of protein detection methods: universal (staining dyes, metal cations, silver, fluorescent, radiolabeling), specific (PTMs), and differential display (2-DE).
Evaluation of method properties: sensitivity, linear dynamic range, reproducibility, cost-efficiency, usability, and MS compatibility.
Emphasis on fluorescent dye-based techniques for protein detection and quantitation.

Main Results:

No single method perfectly fulfills all desired properties for protein detection.
Fluorescent dye-based technologies are currently preferred for most proteomic applications.
Radioactive labeling offers superior sensitivity but is impractical due to cost and safety concerns.

Conclusions:

Fluorescent dye-based protein detection and quantitation offer a practical balance of performance for proteomic analysis.
While radioactive labeling is highly sensitive, its routine use is limited.
The selection of a detection method depends on balancing sensitivity, quantitative accuracy, and practical considerations.