Assay Method Information

Assay Name:  Dose Response Assay for Agonists of 5-Hydroxytryptamine (Serotonin) Receptor Subtype 1A (5HT1A)
Description:  A cell line containing the human 5-HT1a receptor, the promiscuous G-alpha-15 protein (Ga15) as well as the beta-lactamase (BLA) reporter-gene under control of the nuclear factor of activated T-cells (NFAT) promoter was used to measure 5-HT1a agonism. When the 5-HT1a receptor is agonized, transcription of the NFAT-BLA gene occurs via a Ga15 protein coupled signaling cascade. The amount of BLA activity is proportional to the concentration of agonist. BLA activity was measured with a fluorescent BLA substrate. The assay was conducted in 1536-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-CT at a concentration equal to its EC100. A Chinese Hamster Ovary (CHO) cell line stably transfected with human 5HT1a receptor, the nuclear factor of activated T-cell-beta lactamase (NFAT-BLA) reporter construct and the G-alpha-15 promiscuous coupling protein was used.
Affinity data for this assay
 

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