A Protocol for Protein Extraction from Lipid-Rich Plant Tissues Suitable for Electrophoresis

• DOI: 10.1007/978-1-62703-631-3_7
• Publication Date: 2013-10-17T15:52:02Z
• Authors: Agnieszka Zienkie...; Juan David Rejón; Juan de Dios Alché; María Isabel Rodr...; Antonio Jesús Castro
• Keywords: Solubility; Suspensions; Organ Specificity; Olea; Chemical Precipitation; Pollen; Electrophoresis, Gel, Two-Dimensional; Lipids; Filtration; Plant Proteins

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.