- Isolation and Quantification of Plant Extracellular Vesicles.
Isolation and Quantification of Plant Extracellular Vesicles.
Bio-protocol (2017-09-05)
Brian D Rutter, Katie L Rutter, Roger W Innes
PMID34541189
RESUMEN
Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the apoplastic wash of Arabidopsis thaliana leaves. The isolated EVs can be quantified using a fluorometric dye to assess total membrane content.
MATERIALES
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Sigma-Aldrich
Medio para gradiente de densidad OptiPrep™, used for cell and subcellular organelle isolation
Sigma-Aldrich
Murashige and Skoog Basal Salt Mixture (MS), powder, suitable for plant cell culture
Sigma-Aldrich
Cóctel de inhibidores de proteasas, for plant cell and tissue extracts, DMSO solution