Assessment of non-specific binding of Wnt3a to exosomal membranes. Exosomes were isolated from primary cortical neurons (2500000 cells) and incubated with and without (±) Wnt3a (10 nM) in a cell free environment (in SFM) for 8 hours. Following the incubation the exosomes were pelleted by centrifugation at 100000xg for 1 hour and the pellets and the supernatants were subjected to western blotting for the detection of Wnt3a and β-actin as an exosomal marker.
Hooper et al. BMC Neuroscience 2012 13:144 doi:10.1186/1471-2202-13-144