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Total Protein Extraction with TCA-Acetone

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

Abstract

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 

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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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