In situ protein binding assay using Fc-fusion proteins

Abstract

This protocol describes an in situ protein-protein interaction assay between tagged recombinant proteins and cell-surface expressed synaptic proteins. The assay is arguably more sensitive than other traditional protein binding assays such as co-immunoprecipitation and pull-downs and provides a visual readout for binding. This assay has been widely used to determine the dissociation constant of binding of transsynaptic adhesion proteins. The step-wise description in the protocol should facilitate the adoption of this method in other laboratories.