How to Measure Receptor Internalization?

To measure internalization, a therapeutic antibody was labeled with a fluorescent tag, pre-bound to target cells and its translocation from cell surface to cytoplasm was monitored by fluorescence imaging. For the purpose of defining the cytoplasmic compartment, cells were stained with a second fluorescent dye.

What is internalization of a receptor?

Definition: A receptor-mediated endocytosis process that results in the movement of receptors from the plasma membrane to the inside of the cell. The process begins when cell surface receptors are monoubiquitinated following ligand-induced activation.

How is antibody internalization measured?

Flow cytometry has been used extensively to measure internalization of antibodies and receptors. Conventional methods require the removal of a fluorescent signal from the cell surface, usually by washing with an acidic buffer to remove the labeled probe or by quenching with an anti-fluorophore antibody.