| Assay Method Information | |
| | Dose Response Cell Based Assay for Antagonists of the 5-Hydroxytryptamine Receptor Subtype 1E (5HT1E |
| Description: | To measure 5-HT1E activity, a Chinese Hamster Ovary (CHO) cell line was developed at the Scripps Molecular Research Institute Screening center. This line contained the human 5-HT1E receptor, the promiscuous G-alpha-15 protein (Ga15), and the beta-lactamase (BLA) reporter-gene under control of the nuclear factor of activated T-cells (NFAT) promoter. When cells were agonized with a 5HT1E agonist, transcription of the NFAT-BLA gene occurred via the Ga15 protein-coupled signaling cascade. The amount of BLA activity was proportional to the concentration of agonist. BLA activity was measured with a fluorescent BLA substrate. The assay was conducted in 384-well format. Each compound dilution series was assayed in triplicate. All data reported were normalized on a per-plate basis to wells that contained cells in the presence of 5-HT at a concentration equal to its EC90. The CHO cell line was cultured in T-175 sq cm Corning flasks (part 431080) at 37 deg C and 95% RH. The HTS assay began by di |
| Affinity data for this assay | |
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