title: A competitive receptor binding radioassay for beta-1 and beta-2 adrenergic agents.
abstract: A rapid and sensitive competitive receptor binding assay for beta-1 and beta-2 adrenergic binding for adrenergic agents has been developed. The steps that are critical for the success of the assay are given in detail so that the assay can be set up in any routine laboratory with relative ease. The rationale behind the use of specific reagents is discussed. The assay requires microgram quantities of test compound, a radiolabeled specific beta adrenergic antagonist [3H]dihydroalprenolol (DHA), and turkey erythrocyte beta-1 and rat erythrocyte beta-2 receptor membranes. Serial dilutions of sample are incubated with appropriate receptor membranes and DHA for 1 hr at room temperature. After equilibrium is attained, the bound radioligand is separated by rapid filtration under vacuum through Whatman GF/B filters. The amount of bound DHA trapped on the filter is inversely proportional to the degree of beta-1 or beta-2 adrenergic binding of the sample. Separation of bound from free radioligand by filtration permits rapid determination of a large number of samples. This assay quantitates and differentiates beta-1 and beta-2 adrenergic binding of synthetic adrenergic agents.
mesh_terms: D000318:Adrenergic beta-Agonists / Q000032:analysis*; D000818:Animals; D001667:Binding, Competitive; D004082:Dihydroalprenolol / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism*; D007700:Kinetics; D011869:Radioligand Assay / Q000379:methods; D051381:Rats; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D014422:Turkeys
chemical_list: D000318:Adrenergic beta-Agonists; D011943:Receptors, Adrenergic, beta; D004082:Dihydroalprenolol