Problem:

I'm beginning to do some cell-binding assays and I would like for my proteins to not be endocytosed by my mammalian cells. Typical suggestions are for the cells to be kept on ice and that the binding experiment also occurs when the cells are chilled.
However, to be sure it has been suggested that I use endocytosis inhbitors.

Required:

Question 1: When are the endocytosis inhibitors necessary?

Question 2: How would I know that endocytosis inhibitors are necessary?

Question 3: Which ones would I actually use?

I recognize that there are both clathrin and non-clathrin mechanisms of internalization. Which mechanism would be more useful to study for an antibody-receptor interaction?

Please provide some suggestions on endocytosis inhibitors.