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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 Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

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Related Experiment Video

Updated: Jun 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
10:51

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

Protein extraction for 2DE.

Claus Zabel1, Joachim Klose

  • 1Charite - University Medicine Berlin, Institute for Human Genetics, Augustenburger Platz 1, 13353, Berlin, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|April 22, 2009
PubMed
Summary
This summary is machine-generated.

This updated protein extraction protocol for two-dimensional gel electrophoresis (2DE) offers both total and fractionated methods. It ensures protein integrity for proteomics research by minimizing dilution and avoiding harsh steps.

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

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Last Updated: Jun 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
10:51

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

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

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species
08:31

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

Published on: October 15, 2013

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • Two-dimensional gel electrophoresis (2DE) is a cornerstone technique in proteomics.
  • Existing protein extraction protocols require updates to align with current research demands.
  • Over 30 years of experience inform this refined methodology.

Purpose of the Study:

  • To present an updated protein extraction protocol for 2DE.
  • To offer both a rapid total extraction and a specialized fractionated extraction method.
  • To enhance protein recovery and integrity for subsequent proteomic analyses.

Main Methods:

  • Developed a total protein extraction protocol suitable for standard 2DE applications.
  • Developed a fractionated protein extraction protocol for specialized applications (e.g., membrane, nuclear proteins).
  • Emphasized early addition of protease inhibitors to frozen tissue and maintenance of near-physiological pH (7.1) to preserve protein solubility and integrity.

Main Results:

  • The total extraction protocol is fast, simple, and requires no centrifugation.
  • The fractionated protocol addresses the extraction of challenging protein subsets.
  • The protocol avoids precipitation/re-solubilization steps that can lead to protein loss.
  • The extraction method is compatible with advanced 2DE staining and mass spectrometry.

Conclusions:

  • The updated protocol provides a robust and versatile approach for protein extraction in proteomics.
  • It ensures high-quality protein samples for various 2DE applications and downstream analyses.
  • This method supports the analysis of a broader range of proteins, including those from membranes and nuclei.