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Isolation of exosomes from heart and serum
The heart was digested using a protocol, previously described [25]. The heart homogenate was sequentially filtered through a 40 lm mesh filter (BD, Franklin Lakes, NJ, USA) and a 0.2 lm syringe filter (Thermo Scientific, Rockford, IL, USA). The filtrate was passed through a 100 kD cut-off filter (Amicon ultra 15; MIlipore Tullagreen, Carrigtwohill, Co. Cork, Ireland). Exosomes were isolated from the concentrated filtrate by using ultracentrifugation. Briefly, the filtrate was sequentially centrifuged at 300 9 g for 10 min. at 4°C, 2000 9 g for 10 min. at 4°C, and at 10,000 9 g for 30 min. at 4°C to discard cells, membranes and debris. The supernatant was centrifuged at 100,000 9 g for 70 min. at 4°C to pellet exosomes. The exosome pellet was washed with cold PBS and resuspended in 1 ml of cold PBS (Invitrogen, Carlsbad, CA, USA).
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|Archiver|手机版|外泌体之家 | exosomes & microvesicles
发表于 2017-11-3 15:43:20
