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| Solution Information | help | |
| Enzyme: | Sphingosine 1-phosphate receptor 3 | |
| inhibitor: | BDBM75604 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine whether powder samples of compounds identified as active in the assay "Late-stage fluorescence dose-response cell-based assay to identify agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4): Synthesized compounds" (AID 463107) were nonselective agonists as assayed by activation of S1P3. In this assay, a Chinese Hamster Ovary (CHO) cell line containing human S1P3 and the beta-lactamase (BLA) reporter-gene under control of the nuclear factor of activated T-cells (NFAT) promoter was used to measure S1P3 agonism by test compound. Stimulation of S1P3 by S1P induces transcription of NFAT-BLA via a G-alpha16 protein coupled signaling cascade, and an increase in BLA activity. BLA activity is measured using a fluorescent BLA substrate. As designed, a compound that acts as a S1P3 agonist will increase NFAT-BLA transcription and increase well fluorescence. Compounds were tested in triplicate using a 10-point, 1:3 dilution series st | |
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