2D PAGE: Sample Preparation and Fractionation pp 225-239

Part of the Methods in Molecular Biology™ book series (MIMB, volume 424)

Fractionation of Complex Protein Mixtures by Liquid-Phase Isoelectric Focusing

  • Julie Hey
  • Anton Posch
  • Andrew Cohen
  • Ning Liu
  • Adrianna Harbers

Summary

Protein fractionation is essential to uncovering low-abundance proteins in complex protein mixtures. Many common methods and techniques are used to fractionate proteins, including chromatography (size exclusion, affinity, ion exchange, etc.), electrophoresis, and solution chemistry. Regardless of the method employed, the ultimate goal of protein fractionation is to enable more protein analysis by today’s current proteomics technologies, such as one- (1-DGE) or two-dimensional gel electrophoresis (2-DGE) and liquid-chromatography and tandem mass spectrometry (LC-MS/MS).

The MicroRotofor\(^{\rm TM}\) isoelectric focusing (IEF) cell fractionates proteins in free solution according to their isoelectric point (pI). We demonstrate the ability of the MicroRotofor to enrich low-abundance proteins in mouse brain tissue, thus enabling further identification of potential biomarker candidates.

Key Words

Carrier ampholytes 1-D electrophoresis 2-D electrophoresis fractionation liquid phase isoelectric focusing low-abundance proteins mass spectrometry protein purification proteomics Rotofor 

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Julie Hey
    • 1
  • Anton Posch
    • 2
  • Andrew Cohen
    • 1
  • Ning Liu
    • 1
  • Adrianna Harbers
    • 1
  1. 1.Bio-Rad Laboratories, Inc.Hercules
  2. 2.Bio-Rad Laboratories GmbHMunichGermany

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