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Methods Mol Biol. 2012;878:111-20. doi: 10.1007/978-1-61779-854-2_7.

2-DE-based proteomics for the analysis of metastasis-associated proteins.

Author information

1
Department of Molecular and Applied Biosciences, School of Life Sciences, University of Westminster, London, UK. dwekm@wmin.ac.uk

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.

PMID:
22674129
DOI:
10.1007/978-1-61779-854-2_7
[Indexed for MEDLINE]

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