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PEG precipitation

CK Christoph Keysberg
OH Oliver Hertel
LS Louise Schelletter
TB Tobias Busche
CS Chiara Sochart
JK Jörn Kalinowski
RH Raimund Hoffrogge
KO Kerstin Otte
TN Thomas Noll
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Precipitation with polyethylene glycol was carried out as noted by Rider et al. (2016). Culture broth was centrifuged 5 min at 500g and 30 min at 2000g. The precipitation mixture (16% PEG6000 (Merck; Darmstadt, Germany), 1 mM NaCl), was added 1:1 (v/v) to the supernatant, mixed and stored at 4 °C overnight. Precipitated components were sedimented by centrifugation at 4000g for 75 min. The supernatant was thoroughly removed, and the pellet was taken off in PBS and transferred into a 13 ml polypropylene tube (Beckman Coulter; Brea CA, USA) before being centrifuged at 100,000g for 70 min using a TH-641 rotor (Thermo Fisher Scientific; Waltham MA, USA) in an Ultra-Pro 80 centrifuge (Thermo Fisher Scientific; Waltham MA, USA). The exosome-pellet was resuspended in 100 μl PBS with 1 mM PMSF.

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