Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts

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.

Published in

Nature Protocols
2007, Volume: 2, number: 2, pages: 259-262

UKÄ Subject classification

Cell Biology


Publication identifier

DOI: https://doi.org/10.1038/nprot.2007.26

Permanent link to this page (URI)

https://res.slu.se/id/publ/93622