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Sample Preparation for 2D Electrophoresis and CE-Based Proteomics

  • Judit M. Nagy
  • Alexandria Lipka
  • Fiona Pereira
  • Nicky Marlin
  • Stuart HassardEmail author
Chapter

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.

Keywords

Capillary electrophoresis Consistent chemical parameters Proteomics 

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Judit M. Nagy
    • 1
  • Alexandria Lipka
    • 2
  • Fiona Pereira
    • 3
  • Nicky Marlin
    • 4
  • Stuart Hassard
    • 5
    Email author
  1. 1.Department of ChemistryImperial College LondonLondonUK
  2. 2.deltaDOT QSTP-LLCQatar Science & Technology ParkDohaState of Qatar
  3. 3.Department of ChemistryImperial College LondonLondonUK
  4. 4.deltaDOT LtdLondonUK
  5. 5.London BioScience Innovation CentreLondonUK

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