Rabbit anti alix

Exosome-specific markers were detected by western blot analysis (WB) [19 (link),40 (link)]. Thirty micrograms of exosomes were used to perform SDS-PAGE, followed by semidry transfer. The following primary antibodies were used (overnight incubation): rabbit anti-ALIX (Abcam), rabbit anti-Tsg101 (Abcam), rabbit anti-calnexin (Cell Signaling Technology) and mouse anti-Hsp70 (Santa Cruz Biotechnology). The following peroxidase-conjugated antibodies were used: anti-rabbit (Abcam) and anti-mouse (Merck). The membranes were incubated in ECL (Cell Signaling Technology). Proteins were visualized using a ChemiDocTM XRS + system (BioRad). Protein lysate of human adipose MSCs were used as a positive control.

Western blots were realized as previously described [25 (link)], by using the following primary antibodies: mouse anti-CD81 (200 ng/mL, Santa-Cruz, Dallas, TX, USA), mouse anti-CD63 (500 ng/mL, BD Pharmingen, NJ, USA), mouse anti-CD9 (100 ng/mL, Millipore, Burlington, MA, USA), rabbit anti-Tsg101 (1 µg/mL, Abcam, Cambridge, UK), rabbit anti-Alix (1 µg/mL, Abcam), mouse anti-Thy1 (0.5 µg/mL, Biolegend, San Diego, CA, USA), rabbit anti-Tom20 (1/500, Sigma), or goat anti-Calnexin (2 µg/mL, Abcam). After incubation with the secondary antibody, membranes were visualized using the Odyssey Infrared Imaging System (LI-COR Biosciences, Lincoln, NE, USA).