title: A protocol for protein extraction from lipid-rich plant tissues suitable for electrophoresis.
abstract: Plant tissues contain high levels of nonprotein contaminants such as lipids, phenolic compounds, and polysaccharides among others, which interfere with protein extraction and electrophoretic separation. Preparation of good-quality protein extracts is a critical issue for successful electrophoretic analysis. Here, we describe a three-step method for protein extraction from lipid-rich plant tissues, which is suitable for both 1-D and 2-D electrophoresis and is compatible with downstream applications. The protocol includes prefractionation, filtration, and TCA/acetone precipitation steps prior to protein resolubilization.
authors.affiliation: Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas (CSIC), Granada, Spain.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Zienkiewicz
authors.forename: Juan David
authors.initials: JD
authors.lastname: Rejón
authors.forename: Juan
authors.initials: J
authors.lastname: de Dios Alché
authors.forename: María Isabel
authors.initials: MI
authors.lastname: Rodríguez-García
authors.forename: Antonio Jesús
authors.initials: AJ
authors.lastname: Castro
mesh_terms: D011232:Chemical Precipitation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005374:Filtration; D008055:Lipids / Q000737:chemistry*; D031658:Olea / Q000378:metabolism; D009928:Organ Specificity; D010940:Plant Proteins / Q000302:isolation & purification*; D011058:Pollen / Q000378:metabolism; D012995:Solubility; D013535:Suspensions
chemical_list: D008055:Lipids; D010940:Plant Proteins; D013535:Suspensions