Preparation of Plant Protein Samples for 2-D PAGE

  • David W. M. Leung
Part of the Springer Protocols Handbooks book series (SPH)


A critical step in the application of O’Farrell’s (1) two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) in plant biology research is the preparation of plant protein samples free of artefactual protein modifications and without adversely affecting gel resolving power and reproducibility. However, it is important to note that many problems could be encountered in this step. A large part of the plant cell volume is often occupied by the vacuole. Numerous compounds found therein, upon release during disruption of plant cells, might have detrimental or interfering effects on protein extraction. These include organic acids, phenolic compounds, proteases, pigments, polysaccharides, and so on.


Phenolic Compound Protamine Sulfate Page Analysis Pinus Radiata Hypocotyl Segment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biological Chem. 250, 4007–4021.Google Scholar
  2. 2.
    Mayer, J. E., Hahne, G., Palme, K., and Schell, J. (1987) A simple and general plant tissue extraction procedure for two-dimensional gel electrophoresis. Plant Cell Reports 6, 77–81.CrossRefGoogle Scholar
  3. 3.
    De Wit, P. J. G. M. and Spikman, G. (1982) Evidence for the occurrence of race-and cultivar-specific elicitors of necrosis in intercellular fluids of compatible interactions of Cladosporium fulvum and tomato. Physiol. Plant Path. 21, 1–11.CrossRefGoogle Scholar
  4. 4.
    Jeknic, Z. and Chen, T. H. H. (1999) Changes in protein profiles of poplar tissues during the induction of bud dormancy by short-day photoperiods. Plant Cell Physiol. 40, 25–35.Google Scholar
  5. 5.
    Suo Y. and Leung, D. W. M. (2002) Accumulation of extracellular pathogenesis-related proteins in rose leaves following inoculation of in vitro shoots with Diplocarpon rosae. Scientia Hort. 93, 167–178.CrossRefGoogle Scholar
  6. 6.
    Li, M. and Leung, D. W. M. (2001) Protein changes associated with adventitious root formation in hypocotyls of Pinus radiata. Biol. Plant. 44, 33–39.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2005

Authors and Affiliations

  • David W. M. Leung
    • 1
  1. 1.School of Biological Sciences, University of CanterburyNew Zealand

Personalised recommendations