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Sample Preparation for 2D Electrophoresis and CE-Based Proteomics

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Book cover Sample Preparation in Biological Mass Spectrometry

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Abstract

This chapter describes sample preparation for gel-based proteomics, capillary electrophoresis and capillary electrophoresis coupled with mass spectrometry. Two dimensional gel electrophoresis (2D-PAGE) remains one of the most commonly used separation techniques for complex protein mixtures. Stringent and consistent sample preparation is essential for reproducible and high quality 2D-PAGE analysis of complex protein mixtures. The goal of this step is to produce homogeneous sample, free from contaminants and with consistent chemical parameters (pH, salt concentration etc) that will allow the user to generate reproducible results. Capillary electrophoresis (CE) separates analytes from a complex mixture with high resolution based on differential migration through a liquid filled capillary in a strong electric field. CE is easily automated and lends itself well to microfluidic device technology. The evolution of these microfluidic devices allows the potential for the hyphenation of CE to other separation and identification methods like gel chromatography, high performance liquid chromatography (HPLC) and mass spectrometry.

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Authors and Affiliations

  1. Department of Chemistry, Imperial College London, SW7 2AZ, London, UK

    Judit M. Nagy

  2. deltaDOT QSTP-LLC, Qatar Science & Technology Park, Doha, State of Qatar

    Alexandria Lipka

  3. Department of Chemistry, Imperial College London, SW7 2AZ, London, UK

    Fiona Pereira

  4. deltaDOT Ltd, SW7 2BP, London, UK

    Nicky Marlin

  5. London BioScience Innovation Centre, 2 Royal College Street, London, NW1 ONH, UK

    Stuart Hassard

Authors
  1. Judit M. Nagy
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  2. Alexandria Lipka
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  3. Fiona Pereira
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  4. Nicky Marlin
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  5. Stuart Hassard
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Correspondence to Stuart Hassard .

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Editors and Affiliations

  1. , Department of Genetics and Complex Disea, Harvard School of Public Health, Harvard, Huntington Ave., SPH-1 665, Boston, 02115, Massachusetts, USA

    Alexander R. Ivanov

  2. , Research and Development, Pressure BioSciences, Inc., Norfolk Ave. 14, South Easton, 02375, Massachusetts, USA

    Alexander V. Lazarev

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Nagy, J.M., Lipka, A., Pereira, F., Marlin, N., Hassard, S. (2011). Sample Preparation for 2D Electrophoresis and CE-Based Proteomics. In: Ivanov, A., Lazarev, A. (eds) Sample Preparation in Biological Mass Spectrometry. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-0828-0_3

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