Extracellular Vesicle Isolation and Analysis by Western Blotting

Emma J K Kowal; Dmitry Ter-Ovanesyan; Aviv Regev; George M Church

doi:10.1007/978-1-4939-7253-1_12

Extracellular Vesicle Isolation and Analysis by Western Blotting

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

Authors

Emma J K Kowal^{1

2}, Dmitry Ter-Ovanesyan^{3

4

5

6}, Aviv Regev^{7

8}, George M Church^{1

2}

Affiliations

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.

PMID: 28828654
DOI: 10.1007/978-1-4939-7253-1_12

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.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Blotting, Western* / methods
Cell Fractionation* / methods
Extracellular Vesicles* / chemistry
Extracellular Vesicles* / metabolism
Humans

Abstract

Publication types

MeSH terms

Grants and funding