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Evaluating the Efficacy of Subcellular Fractionation of Blast Cells Using Live Cell Labeling and 2D DIGE

  • Yin Ying Ho
  • Megan Penno
  • Michelle Perugini
  • Ian Lewis
  • Peter HoffmannEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 854)

Abstract

Labeling of exposed cell surface proteins of live cells using CyDye DIGE fluor minimal dyes is an efficient strategy for cell surface proteome profiling and quantifying differentially expressed proteins in diseases. Here we describe a strategy to evaluate a two-step detergent-based protein fractionation method using live cell labeling followed by visualization of the fluorescently labeled cell surface proteins and fractionated proteins within a single 2D gel.

Key words

Live cell labeling 2D difference gel electrophoresis Detergent-based fractionation Cell surface proteins Membrane proteins 

References

  1. 1.
    Mayrhofer C, Krieger S, Allmaier G, Kerjaschki D (2006) DIGE compatible labeling of surface proteins on vital cells in vitro and in vivo. Proteomics 6:579–585.PubMedCrossRefGoogle Scholar
  2. 2.
    Strober W (2001) Monitoring cell growth. In: Coligan JE, Bierer BE, Margulies DH, Sherach EM, Strober W (eds). Current Protocols in Immunology, 5th edn. Wiley, New York.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Yin Ying Ho
    • 1
  • Megan Penno
    • 1
  • Michelle Perugini
    • 2
  • Ian Lewis
    • 2
  • Peter Hoffmann
    • 1
    Email author
  1. 1.Adelaide Proteomics CentreUniversity of AdelaideAdelaideAustralia
  2. 2.Division of HaematologySA PathologyAdelaideAustralia

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