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Difference Gel Electrophoresis (DIGE)

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The Protein Protocols Handbook

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

The proteome is a dynamic entity, constantly changing both in levels of protein expression as well as in posttranslational modification and localization, all of which are hidden in the static DNA code. The repertoire of techniques and associated instrumentation which is now being applied to proteomics experiments is expanding exponentially, and although a complete visualization of the proteome is still beyond reach of any single technique, several technology platforms exist that can provide complementary datasets. Proteomics in the clinical setting is also developing, providing a major impact on the way in which diseases will be diagnosed, treated and monitored (1).

In particular, approaches involving two-dimensional (2-D) gel electro-phoresis (2-DE) separations and subsequent protein identification using mass spectrometry (MS) have been useful for imaging thousands of intact proteins (including modified/proteolyzed forms) in a single run, but quantification has been challenging, particularly over a large sample set. Difference gel electro-phoresis (DIGE) has proven to be a powerful quantitative technology for differential-expression proteomics on a global level, where the individual abundance changes for thousands of intact proteins can be simultaneously monitored in replicate samples over multiple variables with statistical confidence (see Note 1). This includes quantitative information on protein isoforms that arise due to a charged post translational modification that results in a change in the isoelectric point of the protein (such as acetylation or phosphorylation), and splice variants or protein processing events, all of which can be resolved for individual quantification and subsequent analysis by mass spectrometry.

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Author information

Authors and Affiliations

  1. Proteomics Laboratory, Mass Spectrometry Research Center, Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA

    David B. Friedman

  2. Cambridge Centre for Proteomics, Cambridge Systems Biology Centre, Department of Biochemistry, University of Cambridge, London, UK

    Kathryn S. Lilley

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  1. David B. Friedman
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  2. Kathryn S. Lilley
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Editors and Affiliations

  1. School of Life Sciences, University of Hertfordshire, Hatfield, Hertfordshire, UK

    John M. Walker

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Friedman, D.B., Lilley, K.S. (2009). Difference Gel Electrophoresis (DIGE). In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-198-7_39

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  • DOI: https://doi.org/10.1007/978-1-59745-198-7_39

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-474-6

  • Online ISBN: 978-1-59745-198-7

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