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Merck

Isolation and Quantification of Plant Extracellular Vesicles.

Bio-protocol (2017-09-05)
Brian D Rutter, Katie L Rutter, Roger W Innes
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.

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Base Trizma®, Primary Standard and Buffer, ≥99.9% (titration), crystalline
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MES hydrate, ≥99.5% (titration)
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Medio para gradiente de densidad OptiPrep, used for cell and subcellular organelle isolation
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Calcium chloride, anhydrous, granular, ≤7.0 mm, ≥93.0%
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Murashige and Skoog Basal Salt Mixture (MS), powder, suitable for plant cell culture
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Cóctel de inhibidores de proteasas, for plant cell and tissue extracts, DMSO solution
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Cloruro de sodio, BioXtra, ≥99.5% (AT)
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Dipropyl disulfide, 98%