Development of a scintiplate assay for recombinant human alpha(2B)-adrenergic receptor.
A high-throughput solid-phase platform for ligand-binding assays using microtiter plates (Scintiplates) has been developed using the scintillation proximity assay principle. The system has been developed using human alpha(2B)-adrenergic receptor (alpha(2B)-AR) expressed from Semliki Forest virus vectors in CHO cells. Alpha(2B)-AR bind natural (adrenaline and noradrenaline) and synthetic ligands with different affinities to mediate a variety of physiological and pharmacological responses. Antagonist radioligands were used for the binding experiments, and the values obtained for the binding constants with the Scintiplate system are in good agreement with those obtained by the traditional filter-binding assay system. The Scintiplate assay offers the advantages of a high-throughput format over the filter-binding assay and is amenable for screening many compounds rapidly for generation of leads.
Institute of Biotechnology (Biocenter 3), University of Helsinki, P.O. Box 65, Viikinkaari 1, FIN-00014, Helsinki, Finland.
S
S
Sen
V-P
VP
Jaakola
H
H
Heimo
P
P
Kivelä
M
M
Scheinin
K
K
Lundstrom
A
A
Goldman
D000818:Animals; D001667:Binding, Competitive; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D005374:Filtration; D006801:Humans; D007700:Kinetics; D008024:Ligands; D011485:Protein Binding; D018341:Receptors, Adrenergic, alpha-2 / Q000032:analysis* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis; D012588:Scintillation Counting / Q000379:methods*
C547771:ADRA2B protein, human; D008024:Ligands; D018341:Receptors, Adrenergic, alpha-2; D011994:Recombinant Proteins