| Assay Method Information | |
| | Detecting inositol monophosphate production |
| Description: | S1P2 Agonist-promoted G-protein responses were determined by measuring changes in intracellular inositol monophosphate using the IP-one TB kit (CisBio, Cat #: 62IPAPEJ) based on time-resolved fluorescence resonance energy transfer (TR-FRET) technology. AssayComplete Cell Plating 2 Reagent was removed and replaced with 1 IP-one stimulation buffer (as per manufacturer's instructions) along with test or control compounds at the desired concentrations. Following a 60-minute incubation at 37 C. and 5% CO2 in a humidified CO2 and temperature-controlled incubator, the components of the IP-one TB kit were added as per the manufacturer's instructions. After an hour incubation at room temperature, plates were analyzed by a BMG PheraStar microplate reader. Responses were measured as the ratio of signal over background, fluorescence emission at 665 nm to fluorescence emission at 620 nm. |
| Affinity data for this assay | |
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