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Methods Mol Biol. 2017;1660:143-152. doi: 10.1007/978-1-4939-7253-1_12.

Extracellular Vesicle Isolation and Analysis by Western Blotting.

Author information

1
Department of Genetics, Harvard Medical School, Boston, MA, USA.
2
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
3
Department of Genetics, Harvard Medical School, Boston, MA, USA. dterovanesyan@fas.harvard.edu.
4
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. dterovanesyan@fas.harvard.edu.
5
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA. dterovanesyan@fas.harvard.edu.
6
Broad Institute of MIT and Harvard, Cambridge, MA, USA. dterovanesyan@fas.harvard.edu.
7
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
8
Department of Biology, MIT, Cambridge, MA, USA.

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.

KEYWORDS:

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

PMID:
28828654
DOI:
10.1007/978-1-4939-7253-1_12
[Indexed for MEDLINE]

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