Analysis of protein-protein interactions using LUMIER assays

  • Methods Mol Biol. 2013:1064:17-27. doi: 10.1007/978-1-62703-601-6_2.
Sonja Blasche  1 ,  Manfred Koegl
Affiliations
  • 1. Genomics and Proteomics Core Facility, German Cancer Research Center, Heidelberg, Germany.
PMID: 23996247 DOI: 10.1007/978-1-62703-601-6_2
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.