A rapid chemical method for lysing Arabidopsis cells for protein analysis

We developed a chemical method for lysing Arabidopsis cells without grinding. In this method, plants are boiled for just 10 minutes in a solution containing a Ca2+ chelator and detergent. Cell extracts prepared by this method were suitable for SDS-PAGE and immunoblot analysis. This method was also applicable to genomic DNA extraction for PCR analysis. Our method was applied to many other plant species, and worked well for some of them.Our method is rapid and economical, and allows many samples to be prepared simultaneously for protein analysis. Our method is useful not only for Arabidopsis research but also research on certain other species.Protein extraction is a frequent procedure in biological research. For preparation of plant cell extracts, it is usually required to grind and homogenize plant materials to physically break the robust cell wall. Sample grinding is laborious and time-consuming, especially when a large number of samples are handled at once. In the case of yeast cells, which also have a cell wall, proteins can be efficiently extracted after cells are treated with alkaline (NaOH) and boiled in SDS-containing solution [1-3]. Although the components and structure of the yeast cell walls [4, for a review] are different from those of the plant cell wall [5, for a review], the simplicity of the yeast method tempted us to seek out a chemical cell-lysis method suitable for plant protein extraction.Here we describe a rapid and simple way of preparing cell extracts from Arabidopsis. We found that in the presence of certain amounts of a Ca2+ chelator and detergent, Arabidopsis cells are quickly lysed just by boiling, without grinding. The method is rapid and economical, and the cell extracts prepared by the method are suitable for SDS-PAGE and immunoblot analysis.In an effort to develop a rapid method of sample preparation, we first simply applied the yeast "alkaline lysis" procedure [3] to Arabidopsis seedlings. Plants were incubated at 100°C for 10 minutes i