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Two-dimensional Gel Electrophoresis

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...

SDS-PAGE

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...

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Related Experiment Video

Updated: May 23, 2026

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

Published on: April 10, 2014

Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional

A A Tan¹, S N Azman, N R Abdul Rani

¹Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia.

Tropical Biomedicine

|March 22, 2012

Summary

A modified TRI-reagent Kit offers superior protein extraction for diverse samples, including cell lines and pathogens. This method enhances protein yield and 2DE gel spot counts for reproducible results.

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Last Updated: May 23, 2026

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

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Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Published on: October 15, 2013

Area of Science:

Proteomics
Biochemistry
Sample Preparation

Background:

Diverse protein sample types necessitate empirical optimization of extraction procedures.
Effective sample preparation is crucial for reproducible 2DE gel electrophoresis, maximizing protein visibility without selective loss.

Purpose of the Study:

To develop a general, reproducible protein extraction method suitable for various sample types.
To compare the efficacy of different protein isolation kits and buffers for diverse biological samples.

Main Methods:

Phenol-chloroform extraction using a modified TRI-reagent Kit.
Comparative analysis of TRI-reagent Kit, EasyBlue Kit, PRO-PREP™ Protein Extraction Solution, and lysis buffer.
Application to diverse samples: MCF-7 cell line, Vibrio cholerae, Cryptocaryon irritans cyst, and liver abscess fat tissue.

Main Results:

The modified TRI-reagent Kit demonstrated the highest protein yield across all tested sample types.
The TRI-reagent Kit method resulted in the greatest number of total protein spots on 2DE gels.
Distinct differences in 2DE gel spot patterns were observed between different extraction methods for each sample type.

Conclusions:

A modified TRI-reagent Kit provides a robust and reproducible method for broad protein extraction from diverse biological sources.
This optimized protocol enhances protein recovery and the number of detectable protein spots in 2DE analysis.
The findings support the empirical determination of optimal protein extraction strategies for specific sample types.