Abstract
BackgroundProtein extraction is a frequent procedure in biological research. For preparation of plant cell extracts, plant materials usually have to be ground and homogenized to physically break the robust cell wall, but this step is laborious and time-consuming when a large number of samples are handled at once.ResultsWe 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.ConclusionsOur 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.
Highlights
Protein extraction is a frequent procedure in biological research
Because having the proper concentrations of sodium dodecyl sulfate (SDS) and NaOH in the solution was critical in the yeast method [1,2,3], we examined the effects of different concentrations of SDS and NaOH on protein extraction from Arabidopsis
The NaOH-free solution worked well, while in preliminary experiments, 2× Laemmli sample buffer [6] failed to extract proteins from unground plants. This suggested that ethylenediaminetetraacetic acid (EDTA) was necessary for efficient protein extraction, and we examined the effects of EDTA on cell wall loosening
Summary
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 applicable to genomic DNA extraction for PCR analysis. Our method was applied to many other plant species, and worked well for some of them
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