High throughput analysis of nuclear receptor-cofactor interactions.
Various assays have been employed to study the nuclear receptor/cofactor interactions. Coimmunoprecipitation protocols, both yeast and mammalian two-hybrid systems, and electrophoretic mobility shift/supershift assays are all commonly used. One of the most useful assays for studying direct protein-protein interactions is the glutathione-S-transferase "pulldown" assay. We have developed a high-throughput version of this assay that utilizes a filter microplate to allow parallel processing of many samples, significantly reducing the time and reagents required for the assay and increasing the sensitivity of the assay for weaker protein-protein interactions.
Goodson
Michael L
ML
Section of Microbiology, Division of Biological Sciences, University of California at Davis, Davis, CA, USA.
Farboud
Behnom
B
Privalsky
Martin L
ML
D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D014176:Protein Biosynthesis; D025941:Protein Interaction Mapping / Q000379:methods; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins