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| | Receptor Binding Studies and GHS-R Ca2+ Flux Assay |
| Description: | Specific binding was determined by incubation of membranes from GHS-R1a transfected CHO-K cells with 125I-His9-ghrelin in the presence of increasing concentrations of compounds to obtain IC50. Fluorescence emissions from 96 wells were measured simultaneously at excitation and emission wavelength of 488 and 520 nm, respectively. During this time, agonist responses in the absence of ghrelin, or the antagonist effects of compounds on ghrelin-stimulated calcium flux were recorded. Sigmoidal curves were fitted and EC50/IC50 values were determined by GraphPad Prism software. Ghrelin shows an EC50 of 0.2 nM in this assay. |
| Affinity data for this assay | |
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