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| Solution Information | help | |
| Enzyme: | Sphingosine 1-phosphate receptor 2 | |
| inhibitor: | BDBM41709 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: A cell line expressing the beta-lactamase (BLA) reporter-gene under control of the cAMP Response Element (CRE) promoter was used to measure activation of the CRE pathway. Under normal conditions, the cells have low CRE basal activity and therefore cells express low BLA levels. Stimulation of the CRE pathway by an agonist induces transcription of the BLA gene in proportion to agonist concentration, resulting in increased BLA activity. 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) and "Counter screen for S1P2 Agonists: Dose Response High Throughput Cell-Based Screen to Identify | |
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