I am using serum depleted media for isolating exosomes from MSC, the depletion time period was for 36h. But the protein concentration estimated by BCA assay was too low.
300g 10min, 2000g 10min, 10000g 30 min
Transfer to UC tubes
100000g 1h 15min two times, last round followed by a pbs wash
pellet resuspended in 1X PBS
ThanksGanga...
I followed exactly the same protocol, but unfortunately the pellet was not visible might be due to low exosome content and the same was revealed by the protein estimation.
whats the starting volume of your conditioned media for isolation..? usually i do with volume not less than 100 ml and a transparent pellet is visible after the first UC step itself
Hi!
Did you managed to isolate the exosomes?
I am doing the same isolation protocol with 40 and 80 ml but no visible pellet and protein concentration was not detectable
@Lilia
I think you should take atleast 200 ml for the pellet to visible. 200-250 ml is sufficient for the exosome isolation in case of ultracentifugation.
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