Extracellular Vesicle Isolation and Analysis by Western Blotting

  • Emma J. K. Kowal
  • Dmitry Ter-OvanesyanEmail author
  • Aviv Regev
  • George M. Church
Part of the Methods in Molecular Biology book series (MIMB, volume 1660)


Extracellular vesicles (EVs) are released by mammalian cells and are thought to be important mediators of intercellular communication. There are many methods for isolating EVs from cell culture media, but the most popular methods involve purification based on ultracentrifugation. Here, we provide a detailed protocol for isolating EVs by differential ultracentrifugation and analyzing EV proteins (such as the tetraspanins CD9, CD63 and CD81) by western blotting.

Key words

Exosomes Extracellular vesicles Exosome isolation Extracellular vesicle isolation Ultracentrifugation Immunoblotting Western blot Tetraspanins CD63 CD81 CD9 



This work was supported by US National Institutes of Health National Human Genome Research Institute grant P50 HG005550.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Emma J. K. Kowal
    • 1
    • 2
  • Dmitry Ter-Ovanesyan
    • 1
    • 2
    • 3
    • 4
    Email author
  • Aviv Regev
    • 4
    • 5
  • George M. Church
    • 1
    • 2
  1. 1.Department of GeneticsHarvard Medical SchoolBostonUSA
  2. 2.Wyss Institute for Biologically Inspired EngineeringHarvard UniversityBostonUSA
  3. 3.Department of Molecular and Cellular BiologyHarvard UniversityCambridgeUSA
  4. 4.Broad Institute of MIT and HarvardCambridgeUSA
  5. 5.Department of BiologyMITCambridgeUSA

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