Skip to main content

Preparation of Plant Protein Samples for 2-D PAGE

  • Protocol
  • 4104 Accesses

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

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.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   259.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   329.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   329.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biological Chem. 250, 4007–4021.

    CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  Google Scholar 

  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.

    CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Leung, D.W.M. (2005). Preparation of Plant Protein Samples for 2-D PAGE. In: Walker, J.M. (eds) The Proteomics Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-890-0:055

Download citation

  • DOI: https://doi.org/10.1385/1-59259-890-0:055

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-343-5

  • Online ISBN: 978-1-59259-890-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics