Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl-derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h.
|Original language||English (US)|
|Number of pages||4|
|State||Published - Mar 2007|
Bibliographical noteFunding Information:
ACKNOWLEDGMENTS This work was supported by the U.S. Department of Energy, Division of Energy Biosciences (Grant DE-FG02-02ER15295 to N.R.).