Detection of angiotensin II type 1 receptor ligands by a cell-based assay.
This work describes a cell-based assay that does not depend on radioactivity or laboratory animals for the detection of ligands of angiotensin II type 1 receptor (AT1R). The assay makes use of stable transfected Chinese hamster ovary cells (CHO-AT1R) expressing the AT1R. A sequential saturation assay principle was used in which receptor binding sites of the CHO-AT1R cells are blocked by the analyte in a concentration-dependent manner. Afterwards, TAMRA-angiotensin II, a fluorescence-labeled ligand, was added to bind to the remaining free binding sites of the receptor. In consequence, the fluorescence signal determined is inversely proportional to the concentration of the analyte.
Griessner
Matthias
M
Fraunhofer Institute for Biomedical Engineering (IBMT), Am Mühlenberg 13, 14476 Potsdam, Germany. matthias.griessner@ibmt.fraunhofer.de
Bröker
Patrick
P
Lehmann
André
A
Ehrentreich-Förster
Eva
E
Bier
Frank F
FF
D000804:Angiotensin II / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003584:Cytological Techniques; D007700:Kinetics; D008024:Ligands; D008163:Luminescent Measurements / Q000379:methods*; D011485:Protein Binding; D044140:Receptor, Angiotensin, Type 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D008024:Ligands; D044140:Receptor, Angiotensin, Type 1; D000804:Angiotensin II