title: A high-throughput ligand competition binding assay for the androgen receptor and other nuclear receptors.
abstract: Standardized, automated ligand-binding assays facilitate evaluation of endocrine activities of environmental chemicals and identification of antagonists of nuclear receptor ligands. Many current assays rely on fluorescently labeled ligands that are significantly different from the native ligands. The authors describe a radiolabeled ligand competition scintillation proximity assay (SPA) for the androgen receptor (AR) using Ni-coated 384-well FlashPlates and liganded AR-LBD protein. This highly reproducible, low-cost assay is well suited for automated high-throughput screening. In addition, the authors show that this assay can be adapted to measure ligand affinities for other nuclear receptors (peroxisome proliferation-activated receptor gamma, thyroid receptors alpha and beta).
authors.affiliation: Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
authors.forename: Clémentine
authors.initials: C
authors.lastname: Féau
authors.forename: Leggy A
authors.initials: LA
authors.lastname: Arnold
authors.forename: Aaron
authors.initials: A
authors.lastname: Kosinski
authors.forename: R Kiplin
authors.initials: RK
authors.lastname: Guy
mesh_terms: D000728:Androgens / Q000032:analysis / Q000378:metabolism*; D001667:Binding, Competitive; D008024:Ligands; D011869:Radioligand Assay / Q000379:methods*; D011944:Receptors, Androgen / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*
chemical_list: D000728:Androgens; D008024:Ligands; D011944:Receptors, Androgen; D018160:Receptors, Cytoplasmic and Nuclear