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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...
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...
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...
Electrophoresis: Overview01:20

Electrophoresis: Overview

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...

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Related Experiment Video

Updated: Jul 5, 2026

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)
08:44

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)

Published on: October 29, 2009

Transverse urea-gradient gel electrophoresis.

D P Goldenberg1

  • 1University of Utah, Salt Lake City, Utah, USA.

Current Protocols in Protein Science
|April 23, 2008
PubMed
Summary

Urea-gradient gel electrophoresis offers a simpler method to study protein unfolding transitions. This technique quantifies unfolding thermodynamics and kinetics, aiding in the characterization of protein conformational properties.

Area of Science:

  • Protein biochemistry
  • Biophysical chemistry
  • Molecular biology

Background:

  • Characterizing protein conformational properties is crucial for understanding protein function.
  • Monitoring protein unfolding transitions induced by heat or chemical denaturants is a key strategy.
  • Traditional spectroscopic methods can be complex for analyzing these transitions.

Purpose of the Study:

  • To present urea-gradient gel electrophoresis as a simpler alternative for analyzing protein unfolding.
  • To demonstrate the utility of this technique for quantitative analysis of unfolding.
  • To highlight its application in identifying protein sample heterogeneity.

Main Methods:

  • Urea-gradient gel electrophoresis was employed to monitor protein unfolding.

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Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting
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Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting

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Last Updated: Jul 5, 2026

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Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
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Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

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Analysis of Mitochondrial Respiratory Chain Complexes in Cultured Human Cells using Blue Native Polyacrylamide Gel Electrophoresis and Immunoblotting
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  • Analysis of gel patterns allowed for estimation of unfolding free energy change.
  • The rate of the unfolding transition was also determined from the gel patterns.

    • Main Results:

    • Urea-gradient gel electrophoresis provides a quantitative assessment of protein unfolding.
    • The technique successfully identified potential covalent or conformational heterogeneity in protein samples.
    • Gel patterns proved sensitive to hydrodynamic volume, net charge, and conformational stability.

    Conclusions:

    • Urea-gradient gel electrophoresis is a valuable and simpler method for studying protein unfolding.
    • This technique facilitates the comparison of different protein forms, such as natural and recombinant proteins.
    • It offers insights into protein conformational stability and sample integrity.