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| Solution Information | help | |
| Enzyme: | Sphingosine 1-phosphate receptor 1 | |
| inhibitor: | BDBM75603 | |
| 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 S1P1. This assay uses Tango S1P1-bla U2OS cells which express the human Endothelial Differentiation Gene 1 (EDG1; S1P1) linked to a GAL4-VP16 transcription factor via a TEV protease site. The cells also express a beta-arrestin/TEV protease fusion protein and a beta-lactamase (BLA) reporter gene under the control of a UAS response element. Stimulation of the S1P1 receptor by agonist causes migration of the fusion protein to the GPCR, and through proteolysis liberates GAL4-VP16 from the receptor. The liberated VP16-GAL4 migrates to the nucleus, where it induces transcription of the BLA gene. BLA expression is monitored by measuring fluorescence r | |
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