title: Analysis of protein-protein interactions using LUMIER assays.
abstract: 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.
authors.affiliation: Genomics and Proteomics Core Facility, German Cancer Research Center, Heidelberg, Germany.
authors.forename: Sonja
authors.initials: S
authors.lastname: Blasche
authors.forename: Manfred
authors.initials: M
authors.lastname: Koegl
mesh_terms: D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*