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Top-Down Proteomics and Comparative 2D-DIGE Analysis

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Difference Gel Electrophoresis

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

Abstract

The combination of large-scale protein separation techniques, sophisticated mass spectrometry, and systems bioinformatics has led to the establishment of proteomics as a distinct discipline within the wider field of protein biochemistry. Both discovery proteomics and targeted proteomics are widely used in biological and biomedical research, whereby the analytical approaches can be broadly divided into proteoform-centric top-down proteomics versus peptide-centric bottom-up proteomics. This chapter outlines the scientific value of top-down proteomics and describes how fluorescence two-dimensional difference gel electrophoresis can be combined with the systematic analysis of crucial post-translational modifications. The concept of on-membrane digestion following the electrophoretic transfer of proteins and the usefulness of comparative two-dimensional immunoblotting are discussed.

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Acknowledgments

Research in the author’s laboratory has been supported by a project grant from the Kathleen Lonsdale Institute for Human Health Research, Maynooth University, and equipment funding under the Research Infrastructure Call 2012 by Science Foundation Ireland (SFI-12/RI/2346/3).

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

    Kay Ohlendieck

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    Kay Ohlendieck

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Ohlendieck, K. (2023). Top-Down Proteomics and Comparative 2D-DIGE Analysis. 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_2

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  • DOI: https://doi.org/10.1007/978-1-0716-2831-7_2

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