Journal
NATURE PROTOCOLS
Volume 2, Issue 2, Pages 259-262Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2007.26
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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.
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