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Isolation and Quantification of Plant Extracellular Vesicles.

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

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.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

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Trizma® Base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
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MES Hydrat, ≥99.5% (titration)
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OptiPrep Dichtegradientenmedium, used for cell and subcellular organelle isolation
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Calciumchlorid, anhydrous, granular, ≤7.0 mm, ≥93.0%
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Murashige und Skoog Basales Salzgemisch (MS), powder, suitable for plant cell culture
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Proteasehemmer-Cocktail, for plant cell and tissue extracts, DMSO solution
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Natriumchlorid, BioXtra, ≥99.5% (AT)
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Dipropyldisulfid, 98%