For a fluorescent ligand undergoing cellular internalization by this pathway, can I assume that I will be able to see cell membrane fluorescence at some point of incubation?
Yes. It is possible to seen the fluorescent ligand on the cell membrane. You can fix cells at different time point and co-stain with membrane protein markers to check the co-localization.
Thanks Christelle, a related question - I have a ligand and I am not sure how it is getting internalized in cells. If I suspect it be internalized in cells via a receptor mediated endocytic pathway but I do not see any membrane binding at 4 degrees at low time points - that proves otherwise?