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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...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
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...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...

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Related Experiment Video

Updated: Jun 12, 2026

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
12:03

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

Published on: February 24, 2011

Gradient SDS Polyacrylamide Gel Electrophoresis.

J M Walker1

  • 1School of Biological and Environmental Sciences, The Hatfield Polytechnic, Hatfield, Hertfordshire, England.

Methods in Molecular Biology (Clifton, N.J.)
|June 1, 2010
PubMed
Summary
This summary is machine-generated.

Gradient polyacrylamide gel electrophoresis offers enhanced protein separation. These gels sharpen protein bands, broaden the molecular weight fractionation range, and improve resolution for closely related proteins compared to linear gels.

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Electrophoretic Separation of Proteins
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Electrophoretic Separation of Proteins

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Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome
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Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Published on: April 2, 2018

Related Experiment Videos

Last Updated: Jun 12, 2026

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
12:03

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

Published on: February 24, 2011

Electrophoretic Separation of Proteins
08:17

Electrophoretic Separation of Proteins

Published on: June 12, 2008

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome
12:34

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Published on: April 2, 2018

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Polyacrylamide gel electrophoresis (PAGE) is a common technique for protein separation.
  • Fixed-concentration gels have limitations in resolving complex protein mixtures.
  • Gradient PAGE utilizes a gradient of increasing acrylamide concentration to improve separation.

Purpose of the Study:

  • To highlight the advantages of gradient polyacrylamide gels over fixed-concentration gels.
  • To explain the mechanism of protein separation in gradient gels.
  • To demonstrate the improved resolution and fractionation range offered by gradient PAGE.

Main Methods:

  • Preparation of gradient polyacrylamide gels.
  • Electrophoresis of protein samples in gradient gels.
  • Analysis of protein banding patterns and resolution.

Main Results:

  • Gradient gels sharpen protein bands by differentially retarding migrating zones.
  • The pore size gradient increases the range of molecular weights fractionated in a single run.
  • Improved separation of proteins with similar molecular weights is achieved in gradient gels.

Conclusions:

  • Gradient polyacrylamide gels provide superior protein separation compared to fixed-concentration gels.
  • The technique enhances band resolution and broadens the applicability for molecular weight fractionation.
  • Gradient PAGE is a valuable tool for analyzing complex protein samples.