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Comparative 3-Sample DIGE Analysis of Skeletal Muscles

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Book cover Difference Gel Electrophoresis

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

Abstract

The skeletal muscle proteome consists of a large number of diverse protein species with a broad and dynamic concentration range. Since mature skeletal muscles are characterized by a specific combination of contractile cells with differing physiological and biochemical properties, it is essential to determine specific differences in the protein composition of fast, slow, and hybrid fibers. Fluorescence two-dimensional gel electrophoresis (DIGE) is a powerful comparative tool to analyze fiber type-specific differences between fast and slow muscles. In this chapter, the application of the DIGE method for the comparative analysis of different subtypes of skeletal muscles is outlined in detail. A standardized proteomic workflow is described, involving sample preparation, protein extraction, differential fluorescence labeling using a 3-dye system, first-dimension isoelectric focusing, second-dimension slab gel electrophoresis, DIGE image analysis, protein digestion, and mass spectrometry.

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Acknowledgements

Research in the author’s laboratory has been supported by project grants from the Irish Higher Education Authority, the Irish Health Research Board, and Muscular Dystrophy Ireland.

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Authors and Affiliations

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

    Kay Ohlendieck

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  1. Kay Ohlendieck
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Correspondence to Kay Ohlendieck .

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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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Ohlendieck, K. (2018). Comparative 3-Sample DIGE Analysis of Skeletal Muscles. 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_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7268-5_9

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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

  • eBook Packages: Springer Protocols

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