Total Protein Extraction with TCA-Acetone

  • Valérie Méchin
  • Catherine Damerval
  • Michel Zivy
Part of the Methods in Molecular Biology book series (MIMB, volume 355)


We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneous precipitation and denaturation with TCA and 2ME in cold acetone. We also describe protein solubilization prior to IEF, either in classical rod gels or in IPGs, using two different solutions. The procedure is easy to carry out. The major caveats are (1) keep samples at low temperature during extraction, and then (2) manage protein samples at about 22 to 25°C to avoid urea precipitation.

Key Words

protein extraction protein solubilization TCA acetone total proteins chaotropes detergents reducing agents plant proteomic 


  1. 1.
    Damerval, C., Zivy, M., Granier, F., and de Vienne, D. (1988) Two-dimensional electrophoresis in plant biology, in Advances in electrophoresis (Chrambach, A., Dunn, M., and Radola, B., eds.), VCH, Weinheim, New York, pp. 265–340.Google Scholar
  2. 2.
    Wu, F. and Wang, M. (1984) Extraction of proteins for sodium dodecyl sulfate polyacrylamide gel electrophoresis from protease-rich plant tissues. Anal. Biochem. 139, 100–103.CrossRefPubMedGoogle Scholar
  3. 3.
    Damerval, C., de Vienne, D., Zivy, M., and Thiellement, H. (1986) Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat seedling proteins. Electrophoresis 7, 52–54.CrossRefGoogle Scholar
  4. 4.
    Saravanan, R. S. and Rose, J. K. (2004) A critical evaluation of sample extraction techniques for enhanced proteomic analysis of recalcitrant plant tissues. Proteomics 4, 2522–2532.CrossRefPubMedGoogle Scholar
  5. 5.
    Carpentier, S. C., Witters, E., Laukens, K., Deckers, P., Swennen, R., and Panis, B. (2005) Preparation of protein extracts from recalcitrant plant tissues: an evaluation of different methods for two-dimensional gel electrophoresis analysis. Proteomics 5, 2497–2507.CrossRefPubMedGoogle Scholar
  6. 6.
    Méchin, V., Consoli, L., Le Guilloux, M., and Damerval, C. (2003) An efficient solubilization buffer for plant proteins focused in immobilized pH gradients. Proteomics 3, 1299–1302.CrossRefPubMedGoogle Scholar
  7. 7.
    Hurkman, W. and Tanaka, C. (1986) Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. Plant Physiol. 81, 802–806.CrossRefPubMedGoogle Scholar
  8. 8.
    Granier, F. (1988) Extraction of plant proteins for two-dimensional electrophoresis. Electrophoresis 9, 712–718.CrossRefPubMedGoogle Scholar
  9. 9.
    Herbert, B. (1999) Advances in protein solubilisation for two-dimensional electrophoresis. Electrophoresis 20, 660–663.CrossRefPubMedGoogle Scholar
  10. 10.
    Colas des Francs, C., Thiellement, H., and de Vienne, D. (1985) Analysis of leaf proteins by two-dimensional gel electrophoresis: protease action as exemplified by ribulose bisphosphate carboxylase/oxygenase degradation and procedure to avoid proteolysis during extraction. Plant Physiol. 78, 178–182.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Valérie Méchin
    • 1
  • Catherine Damerval
    • 2
  • Michel Zivy
    • 2
  1. 1.UMR de Chimie BiologiqueThiverval GrignonFrance
  2. 2.UMR de Génétique Végétale, La Ferme du MoulonGif-sur-YvetteFrance

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