| Solution Information | help | |
| Enzyme: | Sphingosine 1-phosphate receptor 2 | |
| inhibitor: | BDBM41710 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: A cell line stably expressing both S1P2 receptor and the beta-lactamase (BLA) reporter-gene under control of the cAMP Response Element (CRE) promoter was used to measure activation of the S1P2 receptor. Under normal conditions, the cells have low S1P2 activity and low BLA levels. Stimulation of the CRE pathway with an S1P2 agonist induces transcription of the BLA gene in proportion to agonist concentration, resulting in elevated BLA activity. Test compounds that activate S1P will increase CRE activation and BLA reporter gene levels. This increase is monitored by measuring fluorescence resonance energy transfer (FRET) of a cleavable fluorogenic cell permeable BLA substrate. Protocol Summary: The purpose of this assay is to determine dose response curves for 59 purchased compounds that are structurally similar to a hit compound identified by a previous set of experiments entitled, "Dose Response Cell Based Assay for Agonists of the S1P2 Receptor", (PubChem AID 854 | |
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