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

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...
Western Blotting01:15

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

You might also read

Related Articles

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

Sort by
Same author

Overview of Digital Electrophoresis Analysis.

Current protocols·2023
Same author

One-Dimensional Electrophoresis Using Nondenaturing Conditions.

Current protocols in protein science·2018
Same author

Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy.

Current protocols in immunology·2017
Same author

One-dimensional SDS gel electrophoresis of proteins.

Current protocols in protein science·2012
Same author

One-dimensional SDS gel electrophoresis of proteins.

Current protocols in molecular biology·2012
Same author

Quantitation of DNA and RNA with absorption and fluorescence spectroscopy.

Current protocols in neuroscience·2011
Same journal

A Comprehensive Experimental Guide to Studying Cross-Presentation in Dendritic Cells In Vitro.

Current protocols in immunology·2020
Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020
Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020
Same journal

Immunologic Studies in Humans.

Current protocols in immunology·2020
Same journal

Monoclonal Antibodies to Human Cell Surface Antigens.

Current protocols in immunology·2020
Same journal

Protocols for Experimental Sjögren's Syndrome.

Current protocols in immunology·2020
See all related articles

Related Experiment Video

Updated: Jul 5, 2026

Staining Proteins in Gels
10:55

Staining Proteins in Gels

Published on: July 8, 2008

Staining proteins in gels.

Joachim Sasse1, Sean R Gallagher

  • 1Shriners Hospital for Crippled Children, Tampa, Florida, USA.

Current Protocols in Immunology
|April 25, 2008
PubMed
Summary
This summary is machine-generated.

This guide details protein detection in gels using Coomassie blue, silver, or fluorescent stains. Fluorescent staining offers superior sensitivity and is recommended for optimal protein visualization and gel photography.

More Related Videos

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid
07:47

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Published on: August 14, 2009

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels
06:24

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

Published on: April 21, 2019

Related Experiment Videos

Last Updated: Jul 5, 2026

Staining Proteins in Gels
10:55

Staining Proteins in Gels

Published on: July 8, 2008

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid
07:47

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Published on: August 14, 2009

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels
06:24

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

Published on: April 21, 2019

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Accurate protein detection in gels is crucial for biochemical and proteomic analyses.
  • Traditional staining methods like Coomassie blue and silver staining have limitations in sensitivity and dynamic range.
  • Fluorescent stains offer enhanced sensitivity and are increasingly adopted in protein analysis.

Purpose of the Study:

  • To describe standardized protocols for protein detection in polyacrylamide gels.
  • To compare the efficacy of Coomassie blue, silver, and fluorescent staining methods.
  • To provide protocols for gel imaging and photography of stained proteins.

Main Methods:

  • Detailed protocols for Coomassie blue, silver, and fluorescent staining (SYPRO Orange/Red) of proteins in SDS-polyacrylamide gels.
  • Alternate rapid staining procedures for each method.
  • Protocols for protein staining in nondenaturing gels.
  • Support protocols for photographing stained gels, including fluorescently stained proteins.

Main Results:

  • Fluorescent staining demonstrates higher sensitivity compared to Coomassie blue staining.
  • Fluorescent staining sensitivity is comparable to silver staining.
  • Standardized protocols enable reliable protein detection and visualization.

Conclusions:

  • Fluorescent staining is a highly sensitive and effective method for protein detection in gels.
  • The described protocols provide a comprehensive guide for protein staining and imaging.
  • These methods are applicable to both denaturing and nondenaturing gel electrophoresis.