| Solution Information | help | |
| Enzyme: | Sphingosine 1-phosphate receptor 1 | |
| inhibitor: | BDBM44985 | |
| 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 dose response curves for compounds identified as active in a previous set of experiments entitled, "Primary Cell-Based Assay to Identify Agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4)" (PubChem AID 1509), and that confirmed activity in a set of experiments entitled, "Confirmation Cell-Based Assay to Identify Agonists of the Sphingosine 1-Phosphate Receptor 4 (S1P4)" (PubChem AID 1523). This assay also serves as a counterscreen to determine whether these compounds are nonselective agonists due to activation of S1P1. This assay uses Tango S1P1-bla U2OS cells which express S1P1 (EDG1) 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 proteolysi | |
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