Cd63 dynabeads

Exosomes were analyzed for the presence of surface marker CD63 by flow cytometry using magnetic coated CD63-Dynabeads (Thermo Fisher Scientific, Waltham, MA, United States, Cat No. 10606D) as per the manufacturer's instructions. Isolated exosomes were resuspended in 1 × PBS and bound to magnetic-coated CD63-Dynabeads (Thermo Fisher Scientific, Waltham, MA, United States) overnight at 4°C. The following day, the Dynabeads-bound exosomes were incubated with an anti-CD63 mouse antibody (Abcam, Cat No. ab59479) overnight at 4°C along with an appropriate isotype control. After incubation, the beads were washed two times with 1 × PBS and stained with Alexa Flour 488 anti-mouse secondary antibody (detection antibody) for 1 h at 4°C and analyzed by flow cytometry (Beckman Coulter DxFLEX Flow Cytometer).