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. 2021 Oct 1;297(5):101266. doi: 10.1016/j.jbc.2021.101266

Figure 1.

Figure 1

Exosome purification and characterization.A, schematic describing exosome purification procedure. B, size distribution plot of purified exosomes, as determined by NTA. C, immunofluorescence image of purified exosomes, labeled with an Alexa Fluor 647-conjugated anti-CD63 monoclonal antibody, captured using the Particle Metrix PMX-220 ZetaView camera. D, electron micrograph of negative-stained, purified exosomes. Bar, 100 nm. E, immunoblot analysis of equal proportions of 293F cell and exosome lysates using antibodies specific for the exosomal markers CD81, CD9, CD63, E-cadherin (E-cad), N-cadherin (N-cad), GRP78/BiP, calreticulin (CALR), ERGIC-3, GM130, HSP60, and HSP90. The amount and ratio of cell and exosome lysates were selected empirically to show the different enrichment of the CD81, CD9, and CD63 proteins, were kept constant in all immunoblots, by proportion equaled a 10-fold overloading of exosomes relative to cells, and by amount of protein equaled 15 μg cell lysate protein/lane and 0.15 μg exosome lysate protein/lane.