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  • Title: 2-DE-based proteomics for the analysis of metastasis-associated proteins.
    Author: Dwek M, Peiris D.
    Journal: Methods Mol Biol; 2012; 878():111-20. PubMed ID: 22674129.
    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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