Abstract
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) |
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Pages (from-to) | 259-262 |
Number of pages | 4 |
Journal | Nature Protocols |
Volume | 2 |
Issue number | 2 |
DOIs | |
State | Published - Mar 2007 |
Bibliographical note
Funding Information:ACKNOWLEDGMENTS This work was supported by the U.S. Department of Energy, Division of Energy Biosciences (Grant DE-FG02-02ER15295 to N.R.).