title: In vitro muscarinic receptor radioligand-binding assays.
abstract: G-protein-coupled muscarinic receptors (mAChRs), of which there are five subtypes (M(1)-M(5)), are attractive drug targets for a number of disorders. Described in this unit are radioligand-binding assays for defining the selectivity and affinity of chemical agents at the five mAChR subtypes. Detailed methodologies and troubleshooting strategies are provided for saturation-binding studies, to estimate K(D) and B(max) values, and for competition-binding studies to estimate K(i) values. Emphasis is placed on experimental details that are critical for executing a robust and reliable assay.
authors.affiliation: Theravance, South San Francisco, California, USA.
authors.forename: Tod
authors.initials: T
authors.lastname: Steinfeld
authors.forename: M Teresa
authors.initials: MT
authors.lastname: Pulido-Rios
authors.forename: Scott
authors.initials: S
authors.lastname: Armstrong
authors.forename: Sharath S
authors.initials: SS
authors.lastname: Hegde
mesh_terms: D001667:Binding, Competitive / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D008024:Ligands; D018721:Muscarinic Agonists / Q000494:pharmacology*; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D011869:Radioligand Assay / Q000379:methods*; D011976:Receptors, Muscarinic / Q000378:metabolism*
chemical_list: D008024:Ligands; D018721:Muscarinic Agonists; D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic