Fig. 5.

Alternative rapid isolation techniques of exosomes from concentrated media. (a) Size distribution and EM images of particles isolated from precipitation methods and qEV SEC columns, size bar=200 nm. (b) Precipitation methods isolate significantly more particles (<100 nm) compared to SEC and density gradient purification. (c) Concentration of particles expressed as a ratio per microgram of protein. Both SEC and DG provide superior purity as illustrated by significantly more particles per microgram of protein compared to precipitation protocols. (d) No difference was observed in the particle size composition of different isolation methods. (e) Western blot analysis of 10 µg of protein from each protocol. Exosome-positive markers were enriched in qEV and DG lysates compared to precipitation isolations, and all isolation techniques were absent for Calnexin, which was present only in the cell lystate fraction. n=3±SEM, *p<0.05, **p<0.01, ***p<0.001. CL: cell lysate; EQ: ExoQuick™; ES: Exo-spin™; qEV: size exclusion columns; DG: density gradient.