A simple and general plant tissue extraction procedure for two-dimensional gel electrophoresis

A fast and simple extraction procedure of plant tissue for two-dimensional gel electrophoresis is presented. The procedure is especially useful for the extraction of plant cell suspension cultures, callus and other plant tissues having a high content of phenol oxidases, polysaccharides, polynucleotides, terpenoids and other substances interfering with isoelectric focusing. Due to the speed of the extraction procedure (about 20 min), large numbers of samples containing only milligram amounts of tissue can be easily processed. The simplicity of the method makes it particularly suitable for the extraction of radiolabeled tissues (³⁵S, ³²P). This method is perfectly compatible with silver staining, autoradiography and Western blotting analysis.Abbreviations DTT dithiothreitol - IEF isoelectric focusing - SDS sodium dodecyl sulfate - TEMED N,N,N,N-tetramethylethylenediamine