Analysis of ProteinProtein Interactions Using LUMIER Assays

Co-affinity purification methods can test whether two proteins physically engage in a complex. The assay principle is to enrich cellular extracts for a first protein by a purification step, and then test if a second protein is enriched as well. This principle has been optimized for use at high-throughput in LUMIER (luminescence-based mammalian interactome mapping) assays, which use luciferase-tags for the detection of the second proteins. This protocol is ideal for miniaturization of the assay on microtiter plates and supports a throughput of several hundred interaction tests per week.