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2-DE-Based Proteomics for the Analysis of Metastasis-Associated Proteins

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Metastasis Research Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 878))

Abstract

Two-dimensional electrophoresis (2-DE) is a high-resolution technique for analysis and comparison of complex protein mixtures. With the advent of recent technical developments, its application has become significant in a wide range of fields. This chapter describes a proteomic approach for the analysis of metastasis-associated proteins using pre-fractionation of glycosylated proteins via lectin (HPA) affinity chromatography prior to separation by 2-DE. Guidelines for the preparation and storage of buffers, experimental conditions and protocols of affinity chromatography, isoelectric focussing, and SDS-PAGE conditions are provided. Critical parameters associated with the different steps of 2-DE are discussed.

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Authors and Affiliations

  1. Department of Molecular and Applied Biosciences, School of Life Sciences, University of Westminster, London, UK

    Miriam Dwek & Diluka Peiris

Authors
  1. Miriam Dwek
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  2. Diluka Peiris
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Corresponding author

Correspondence to Miriam Dwek .

Editor information

Editors and Affiliations

  1. School of Life Sciences, Molecular and Applied Biosciences, University of Westminster, New Cavendish Street 115, London, W1W 6UW, United Kingdom

    Miriam Dwek

  2. School of Life Sciences, Oxford Brookes University, Gipsy Lane, Oxford, OX3 0BP, United Kingdom

    Susan A. Brooks

  3. Inst. Anatomie und Experimentelle, Morphologie, Universitätsklinikum Hamburg, Martinistr. 52, Hamburg, 20246, Germany

    Udo Schumacher

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Dwek, M., Peiris, D. (2012). 2-DE-Based Proteomics for the Analysis of Metastasis-Associated Proteins. In: Dwek, M., Brooks, S., Schumacher, U. (eds) Metastasis Research Protocols. Methods in Molecular Biology, vol 878. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-854-2_7

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  • DOI: https://doi.org/10.1007/978-1-61779-854-2_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-853-5

  • Online ISBN: 978-1-61779-854-2

  • eBook Packages: Springer Protocols

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