title: Sample extraction techniques for enhanced proteomic analysis of plant tissues.
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.
authors.affiliation: Department of Plant Biology, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Tal
authors.initials: T
authors.lastname: Isaacson
authors.forename: Cynthia M B
authors.initials: CM
authors.lastname: Damasceno
authors.forename: Ramu S
authors.initials: RS
authors.lastname: Saravanan
authors.forename: Yonghua
authors.initials: Y
authors.lastname: He
authors.forename: Carmen
authors.initials: C
authors.lastname: Catalá
authors.forename: Montserrat
authors.initials: M
authors.lastname: Saladié
authors.forename: Jocelyn K C
authors.initials: JK
authors.lastname: Rose
mesh_terms: D010940:Plant Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D010944:Plants / Q000737:chemistry*; D020543:Proteome / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*
chemical_list: D010940:Plant Proteins; D020543:Proteome