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DIGE Analysis of Clinical Specimens

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Difference Gel Electrophoresis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2596))

Abstract

Two-dimensional difference gel electrophoresis (2D-DIGE) is an elegant gel electrophoretic analytical tool for comparative protein assessment. It is based on two-dimensional gel electrophoresis (2D-GE) separation of fluorescently labeled protein extracts. The tagging procedures are designed to not interfere with the chemical properties of proteins with respect to their pI and electrophoretic mobility, once a proper labeling protocol is followed. The use of an internal pooled standard makes 2D-DIGE a highly accurate quantitative method enabling multiple protein samples to be separated on the same two-dimensional gel. Technical limitations of this technique (i.e., underrating of low abundant, high molecular mass and integral membrane proteins) are counterbalanced by the incomparable separation power which allows proteoforms and unknown PTM (posttranslational modification) identification. Moreover, the image matching and cross-gel statistical analysis generates robust quantitative results making data validation by independent technologies successful.

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Acknowledgments

This work was supported by the Italian Ministry of University and Scientific Research (grant PRIN 201742SBXA to D.C. and PRIN 2015FBNB5Y to C.G.), Fondazione Cariplo (grant 2017-0622 to C.G.), and Agenzia Spaziale Italiana (grant 2018-9-U.O STOPBROS to C.G.).

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Authors and Affiliations

  1. Department of Biomedical Sciences for Health, University of Milan, Segrate, Italy

    Cecilia Gelfi & Daniele Capitanio

  2. IRCCS Istituto Ortopedico Galeazzi, Milan, Italy

    Cecilia Gelfi

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  1. Cecilia Gelfi
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  2. Daniele Capitanio
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Correspondence to Daniele Capitanio .

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  1. Maynooth University, MAYNOOTH, Ireland

    Kay Ohlendieck

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Gelfi, C., Capitanio, D. (2023). DIGE Analysis of Clinical Specimens. In: Ohlendieck, K. (eds) Difference Gel Electrophoresis. Methods in Molecular Biology, vol 2596. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2831-7_14

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  • DOI: https://doi.org/10.1007/978-1-0716-2831-7_14

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