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High-Resolution 2DE

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Two-Dimensional Electrophoresis Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 519))

Summary

About 30 years ago two-dimensional gel electrophoresis (2DE) was developed independently by Klose and O’Farrell representing the combination of two orthogonal separation techniques. In the first dimension the proteins are separated by isoelectric focusing (IEF) according to their isoelectric point. In the second dimension proteins are separated according to their electrophoretic mobility by conventional SDS-PAGE. For IEF two different techniques, immobilized pH gradient (IPG) and carrier-ampholyte-based IEF (CA-based IEF), respectively, are currently applied. With a resolution of up to 10,000 protein spots in one gel, 2DE offers a huge potential to give a comprehensive overview of the proteins present in the examined system. In combination with image analysis and mass spectrometry 2DE is still the method of choice to analyse complex protein samples.

In this chapter we provide detailed protocols for both 2DE systems and give an overview about the latest developments including the two-dimensional difference gel electrophoresis (DIGE) system.

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Acknowledgment

This work was supported by the Bundesministerium fĂĽr Bildung und Forschung (NGFN, FZ 031U119 and 01GR0440) and the Ministerium fĂĽr Wissenschaft und Forschung Nordrhein Westfalen.

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Authors and Affiliations

  1. Medizinisches Proteom-Center, Ruhr-Universitaet Bochum, Universitaetsstr. 150, D-44780, Bochum, Germany

    Katrin Marcus, Cornelia Joppich, Caroline May, Kathy Pfeiffer, Barbara Sitek, Helmut Meyer & Kai Stuehler

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  1. Katrin Marcus
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  2. Cornelia Joppich
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  3. Caroline May
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  4. Kathy Pfeiffer
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  5. Barbara Sitek
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  6. Helmut Meyer
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  7. Kai Stuehler
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Correspondence to Katrin Marcus .

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    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

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    Marcus, K. et al. (2009). High-Resolution 2DE. In: Tyther, R., Sheehan, D. (eds) Two-Dimensional Electrophoresis Protocols. Methods in Molecular Biology, vol 519. Humana Press. https://doi.org/10.1007/978-1-59745-281-6_14

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    • DOI: https://doi.org/10.1007/978-1-59745-281-6_14

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    • Publisher Name: Humana Press

    • Print ISBN: 978-1-58829-937-6

    • Online ISBN: 978-1-59745-281-6

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