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Related Concept Videos

SDS-PAGE01:27

SDS-PAGE

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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...
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Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

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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...
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DNA Agarose Gel Electrophoresis02:35

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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.
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In cloning experiments, both the insert and vector DNA...
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Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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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,...
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Electrophoresis: Overview01:20

Electrophoresis: Overview

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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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Southern Blot02:57

Southern Blot

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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.
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Updated: Jan 2, 2026

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture
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Tricine-SDS-PAGE.

Hermann Schägger1

  • 1Molekulare Bioenergetik, Zentrum der Biologischen Chemie, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, Haus 26, D-60590 Frankfurt, Germany. schagger@zbc.kgu.de

Nature Protocols
|April 5, 2007
PubMed
Summary
This summary is machine-generated.

Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) effectively separates proteins up to 100 kDa, especially those under 30 kDa. This versatile protocol enhances protein identification through improved electroblotting and staining methods.

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Area of Science:

  • Biochemistry
  • Proteomics
  • Molecular Biology

Background:

  • Tricine-SDS-PAGE is a standard technique for protein separation within the 1-100 kDa mass range.
  • It excels at resolving small proteins (<30 kDa) due to lower acrylamide concentrations.
  • Lower acrylamide concentrations facilitate efficient electroblotting, crucial for hydrophobic proteins.

Purpose of the Study:

  • To present a refined protocol for Tricine-SDS-PAGE.
  • To enhance the versatility of Tricine-SDS-PAGE through optimized staining and electroblotting methods.
  • To improve the resolution of hydrophobic proteins and facilitate their identification.

Main Methods:

  • Implementation of a Tricine-SDS-PAGE protocol.
  • Incorporation of efficient Coomassie blue and silver staining techniques.
  • Integration of optimized electroblotting procedures.
  • Application in doubled SDS-PAGE (dSDS-PAGE) and after native PAGE (BN-PAGE, CN-PAGE).

Main Results:

  • The protocol allows for effective separation of proteins in the 1-100 kDa range, with superior resolution for proteins <30 kDa.
  • Enhanced electroblotting efficiency, particularly for hydrophobic proteins.
  • Versatile application with efficient Coomassie blue and silver staining.
  • Successful integration with dSDS-PAGE, BN-PAGE, and CN-PAGE for complex proteomic analyses.

Conclusions:

  • The described Tricine-SDS-PAGE protocol offers a versatile and efficient method for protein separation and analysis.
  • It provides significant advantages for the resolution and identification of small and hydrophobic proteins.
  • The protocol is suitable for various proteomic applications, including advanced techniques like dSDS-PAGE.