title: An ultrahigh-throughput screening assay for estrogen receptor ligands.
abstract: Members of the steroid and thyroid hormone receptor superfamily (nuclear receptors) play diverse roles in mammalian physiology, in both normal and pathological states. For this reason, and because nuclear receptors are natural receptors for lipophilic small molecules, they are important therapeutic targets for the pharmaceutical industry. Here we describe a method for screening for ligands for one of these receptors, the estrogen receptor. The method is rapid, robust, and reliable, and has been used in an ultrahigh-throughput robotic screen. The receptor is crosslinked to a scintillant-containing solid support (FlashPlate) via a receptor-specific antibody. Test compounds are assayed for competition with a radiolabeled estrogen for binding to the immobilized receptor. Receptor-ligand complexes are allowed to form and receptor-bound radioactivity is detected in a scintillation counter. The assay has been designed for both isoforms of the estrogen receptor, alpha and beta, using separate antibodies for each. Given a radioactive tracer and an appropriate antibody, many of which are now commercially available, the assay could be established for any nuclear receptor.
authors.affiliation: Department of Reproductive Therapeutics, R. W. Johnson Pharmaceutical Research Institute, Raritan, New Jersey 08869, USA. gallan@prius.jnj.com
authors.forename: G F
authors.initials: GF
authors.lastname: Allan
authors.forename: A
authors.initials: A
authors.lastname: Hutchins
authors.forename: J
authors.initials: J
authors.lastname: Clancy
mesh_terms: D008024:Ligands; D011960:Receptors, Estrogen / Q000378:metabolism*; D015203:Reproducibility of Results; D012588:Scintillation Counting / Q000379:methods*
chemical_list: D008024:Ligands; D011960:Receptors, Estrogen