Solubilization of Plant Membrane Proteins for Analysis by Two-Dimensional Gel Electrophoresis

摘要: A plasma membrane-enriched fraction prepared from barley roots was analyzed by two-dimensional gel electrophoresis. Four methods of sample solubilization were assessed on silver stained gels. When membranes solubilized with 2% sodium dodecyl sulfate followed addition Nonidet P-40, gels had high background staining and few proteins because incomplete solubilization. Gels in urea P-40 a greater number but molecular weights than 85,000 absent low diffuse. High weight present 4% acetone precipitation streaking remained problem. the best quality obtained when membrane extracted phenol precipitated ammonium acetate methanol; diminished clearly resolved. This method makes possible resolution required for meaningful qualitative quantitative comparisons protein patterns plant proteins.