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

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...
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...
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...

You might also read

Related Articles

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

Sort by
Same author

Evaluation of a Publicly Available Artificial Intelligence Model for Interpretation of Stains and Smears for Infectious Diseases Diagnosis.

The journal of applied laboratory medicine·2026
Same author

You get what you need: Implementation of electronic remote blood refrigerators to reduce inefficiencies in blood product allocation and massive transfusion protocol overactivation.

American journal of clinical pathology·2026
Same author

Retooling the Massive Transfusion Protocol at a Veterans Affairs Medical Center.

Military medicine·2025
Same author

A protocol for transposon insertion sequencing in <i>Schizosaccharomyces pombe</i> to identify factors that maintain heterochromatin.

STAR protocols·2021
Same author

Non-canonical Targets of HIF1a Impair Oligodendrocyte Progenitor Cell Function.

Cell stem cell·2020
Same author

Suppression of proteolipid protein rescues Pelizaeus-Merzbacher disease.

Nature·2020
Same journal

Kinship Inferences for Second-Degree Relatives With a Combination of STRs and Microhaplotypes.

Electrophoresis·2026
Same journal

Optimisation of Electrokinetic Extraction System: Colourimetric Determination of Copper (II) in Sand Using Polymer Inclusion Membrane.

Electrophoresis·2026
Same journal

Novel Phloroglucinol Derivatives as Neuraminidase Inhibitors Identified From Humulus lupulus L. Extract by At-Line Nanofractionation Platform.

Electrophoresis·2026
Same journal

Protein-Based High-Performance Liquid Chromatography and Cyclodextrin-Capillary Electrokinetic Chromatography for the Chiral Separation of Azoles.

Electrophoresis·2026
Same journal

Dynamics of Heparin Translocations Through Solid-State Nanopores.

Electrophoresis·2026
Same journal

Production of Protein Hydrolysates and Bioactive Peptides From Lablab purpureus and Macrotyloma uniflorum via Optimized Extraction and Proteolysis Protocols.

Electrophoresis·2026
See all related articles

Related Experiment Video

Updated: Jun 16, 2026

Preparation of DNA-crosslinked Polyacrylamide Hydrogels
09:06

Preparation of DNA-crosslinked Polyacrylamide Hydrogels

Published on: August 27, 2014

Stabilizing labile DNA-protein complexes in polyacrylamide gels.

Nina Y Sidorova1, Stevephen Hung, Donald C Rau

  • 1Laboratory of Physical and Structural Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. sidorova@mail.nih.gov

Electrophoresis
|January 29, 2010
PubMed
Summary
This summary is machine-generated.

This study enhances the electrophoretic mobility-shift assay (EMSA) by adding triethylene glycol to gels. This stabilizes labile DNA-protein complexes, improving quantitation for a wider range of binding affinities.

More Related Videos

Multimer-PAGE: A Method for Capturing and Resolving Protein Complexes in Biological Samples
07:40

Multimer-PAGE: A Method for Capturing and Resolving Protein Complexes in Biological Samples

Published on: May 5, 2017

Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers
10:17

Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers

Published on: October 5, 2017

Related Experiment Videos

Last Updated: Jun 16, 2026

Preparation of DNA-crosslinked Polyacrylamide Hydrogels
09:06

Preparation of DNA-crosslinked Polyacrylamide Hydrogels

Published on: August 27, 2014

Multimer-PAGE: A Method for Capturing and Resolving Protein Complexes in Biological Samples
07:40

Multimer-PAGE: A Method for Capturing and Resolving Protein Complexes in Biological Samples

Published on: May 5, 2017

Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers
10:17

Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers

Published on: October 5, 2017

Area of Science:

  • Molecular Biology
  • Biochemistry

Background:

  • Electrophoretic mobility-shift assay (EMSA) is crucial for studying DNA-protein interactions.
  • Standard EMSA is limited to strong DNA-protein complexes (K(a)>10(9) M(-1)) due to dissociation in gel matrices.
  • Labile complexes often yield smeared bands, hindering reliable quantitation.

Purpose of the Study:

  • To improve the sensitivity and range of EMSA for studying DNA-protein interactions.
  • To stabilize weak or labile DNA-protein complexes during electrophoretic separation.
  • To enable reliable quantitation of binding for a broader spectrum of molecular interactions.

Main Methods:

  • Incorporation of the osmolyte triethylene glycol into polyacrylamide gel matrices for EMSA.
  • Utilizing restriction endonuclease EcoRI and nonspecific DNA sequences to test stabilization.
  • Comparison of binding quantitation with and without triethylene glycol in EMSA.

Main Results:

  • Addition of triethylene glycol dramatically stabilized labile restriction endonuclease EcoRI complexes with nonspecific DNA.
  • The modified EMSA enabled reliable quantitation of binding for previously unstable complexes.
  • The technique's effective range for investigating protein-DNA complex binding constants was significantly extended.

Conclusions:

  • Triethylene glycol in EMSA gels substantially enhances the stability of DNA-protein complexes.
  • This modification broadens the applicability of EMSA to a wider range of binding affinities.
  • The approach is adaptable to other separation techniques like gel chromatography and capillary electrophoresis (CE).