Protocol abstract
Nature Protocols 1, - 769 - 774 (2006)
Published online: 13 July 2006 | doi:10.1038/nprot.2006.102
Subject Categories: Biochemistry and protein analysis | Isolation, purification and separation | Plant biology
Sample extraction techniques for enhanced proteomic analysis of plant tissues
Tal Isaacson1, Cynthia M B Damasceno1, Ramu S Saravanan1, Yonghua He1, Carmen Catalá1, Montserrat Saladié1 & Jocelyn K C Rose1
Abstract
Major improvements in proteomic techniques in recent years have led to an increase in their application in all biological fields, including plant sciences. For all proteomic approaches, protein extraction and sample preparation are of utmost importance for optimal results; however, extraction of proteins from plant tissues represents a great challenge. Plant tissues usually contain relatively low amounts of proteins and high concentrations of proteases and compounds that potentially can limit tissue disintegration and interfere with subsequent protein separation and identification. An effective protein extraction protocol must also be adaptable to the great variation in the sets of secondary metabolites and potentially contaminating compounds that occurs between tissues (e.g., leaves, roots, fruit, seeds and stems) and between species. Here we present two basic protein extraction protocols that have successfully been used with diverse plant tissues, including recalcitrant tissues. The first method is based on phenol extraction coupled with ammonium acetate precipitation, and the second is based on trichloroacetic acid (TCA) precipitation. Both extraction protocols can be completed within 2 d.
- 228 Plant Sciences Building, Department of Plant Biology, Cornell University, Ithaca, New York 14853, USA.
Correspondence to: Jocelyn K C Rose1 e-mail: jr286@cornell.edu
