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Extracellular Vesicle Isolation and Analysis by Western Blotting

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Extracellular Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1660))

Abstract

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.

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References

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Acknowledgment

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

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Author notes

  1. Emma J.K. Kowal and Dmitry Ter-Ovanesyan contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, Boston, MA, USA

    Emma J. K. Kowal, Dmitry Ter-Ovanesyan & George M. Church

  2. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA

    Emma J. K. Kowal, Dmitry Ter-Ovanesyan & George M. Church

  3. Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA

    Dmitry Ter-Ovanesyan

  4. Broad Institute of MIT and Harvard, Cambridge, MA, USA

    Dmitry Ter-Ovanesyan & Aviv Regev

  5. Department of Biology, MIT, Cambridge, MA, USA

    Aviv Regev

Authors
  1. Emma J. K. Kowal
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  2. Dmitry Ter-Ovanesyan
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  3. Aviv Regev
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  4. George M. Church
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Corresponding author

Correspondence to Dmitry Ter-Ovanesyan .

Editor information

Editors and Affiliations

  1. CloudHealth Genomics, Ltd., Shanghai, China

    Winston Patrick Kuo

  2. Predicine, Inc., Hayward, California, USA

    Shidong Jia

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© 2017 Springer Science+Business Media LLC

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Kowal, E.J.K., Ter-Ovanesyan, D., Regev, A., Church, G.M. (2017). Extracellular Vesicle Isolation and Analysis by Western Blotting. In: Kuo, W., Jia, S. (eds) Extracellular Vesicles. Methods in Molecular Biology, vol 1660. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7253-1_12

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  • DOI: https://doi.org/10.1007/978-1-4939-7253-1_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7251-7

  • Online ISBN: 978-1-4939-7253-1

  • eBook Packages: Springer Protocols

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