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DIGE Analysis of Human Tissues

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Difference Gel Electrophoresis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1664))

Abstract

Two-dimensional difference gel electrophoresis (2-D DIGE) is an advanced and elegant gel electrophoretic analytical tool for comparative protein assessment. It is based on two-dimensional gel electrophoresis (2-DE) 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 two-dye or three-dye systems can be adopted and their choice depends on specific applications. Furthermore, the use of an internal pooled standard makes 2-D DIGE a highly accurate quantitative method enabling multiple protein samples to be separated on the same two-dimensional gel. The image matching and cross-gel statistical analysis generates robust quantitative results making data validation by independent technologies successful.

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Acknowledgements

This work was supported by the Telethon foundation (grant N. GGP08107D to C.G.), EU community (grant BIO-NMD N. 241665 to C.G.) and Italian Ministry of University and Scientific Research (grant FIRB RBRN07BMCT to C.G. and PRIN 2015FBNB5Y).

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Authors and Affiliations

  1. Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, via f.lli Cervi, 93, 20090, Segrate, Milan, Italy

    Cecilia Gelfi & Daniele Capitanio

  2. U.O. Proteomica clinica, IRCCS Policlinico San Donato, 20097, San Donato, Milan, Italy

    Cecilia Gelfi

  3. Istituto di Bioimmagini e Fisiologia Molecolare, CNR, 20090, Segrate, Milan, Italy

    Cecilia Gelfi

Authors
  1. Cecilia Gelfi
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  2. Daniele Capitanio
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Corresponding author

Correspondence to Cecilia Gelfi .

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Editors and Affiliations

  1. Department of Biology, Maynooth University, National University of Ireland, Maynooth, Co. Kildare, Ireland

    Kay Ohlendieck

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Gelfi, C., Capitanio, D. (2018). DIGE Analysis of Human Tissues. In: Ohlendieck, K. (eds) Difference Gel Electrophoresis. Methods in Molecular Biology, vol 1664. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7268-5_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7268-5_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7267-8

  • Online ISBN: 978-1-4939-7268-5

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