Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts

Stéphanie Robert, Jan Zouhar, Clay Carter, Natasha Raikhel

Research output: Contribution to journalArticle

57 Scopus citations

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 languageEnglish (US)
Pages (from-to)259-262
Number of pages4
JournalNature Protocols
Volume2
Issue number2
DOIs
StatePublished - Mar 1 2007

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