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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...
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...
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...
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,...
Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...
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...

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Updated: Jun 22, 2026

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

High-resolution two-dimensional electrophoresis.

Walter Weiss1, Angelika Görg

  • 1Technische Universität München, Fachgebiet Proteomik, Am Forum 2, D-85350 Freising-Weihenstephan, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|June 23, 2009
PubMed
Summary
This summary is machine-generated.

Two-dimensional gel electrophoresis (2-DE) with immobilized pH gradients (IPGs) is a powerful proteomic tool for analyzing complex protein mixtures. This method offers high-resolution protein expression profiling, detecting thousands of proteins simultaneously.

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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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Use of Two Dimensional Semi-denaturing Detergent Agarose Gel Electrophoresis to Confirm Size Heterogeneity of Amyloid or Amyloid-like Fibers
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Use of Two Dimensional Semi-denaturing Detergent Agarose Gel Electrophoresis to Confirm Size Heterogeneity of Amyloid or Amyloid-like Fibers

Published on: April 26, 2018

Area of Science:

  • Proteomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry is a standard technique for proteome projects.
  • While newer methods exist, 2-DE excels in routine parallel expression profiling of complex protein mixtures.
  • Modern 2-DE with immobilized pH gradients (IPGs) has improved reproducibility and separation capabilities.

Purpose of the Study:

  • To provide a comprehensive protocol for high-resolution 2-DE with IPGs for proteome analysis.
  • To detail the steps involved in sample preparation, protein separation, and quantification.
  • To address common challenges and offer solutions for successful proteomic analysis.

Main Methods:

  • High-resolution 2-DE utilizing immobilized pH gradients (IPGs).
  • Protein identification via mass spectrometry.
  • Detailed protocol covering sample preparation, solubilization, isoelectric focusing, equilibration, and SDS-PAGE.
  • Fluorescent dye technologies for protein quantitation.

Main Results:

  • 2-DE can resolve up to 5,000 proteins, routinely analyzing approximately 2,000.
  • The technique can detect and quantify proteins at levels below 1 ng per spot.
  • 2-DE provides a comprehensive protein map reflecting expression levels, isoforms, and post-translational modifications.

Conclusions:

  • High-resolution 2-DE with IPGs remains a robust and essential technology for proteome analysis.
  • The described protocol facilitates detailed protein expression profiling and quantification.
  • Ongoing advancements aim to further enhance the separation of challenging proteins and improve detection sensitivity.