Anti-TfR antibody (5E9C11) was conjugated with EMG 508 ferrofluid. HeLa cells were loaded with TfR antibody-ferrofluid conjugates and internalized into cells in starvation conditions. After TfR antibody-ferrofluid conjugates were delivered to the recycling endosome, cells were mechanically disrupted, ferrofluid-TfR antibody-containing membranes were magnetically isolated and then analyzed by western blot. Bound and unbound fractions (cytosol and unbound membrane) were processed for SDS-PAGE. RAB11A, LC3, and GAPDH were labeled to test the specificity of the two fractions.
