MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT S1P1 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human S1P1 cDNA (GenBank Accession Number NM_001400) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM/F12, 10% FBS, 10 μg/mL puromycin
Stable in culture for minimum of two months
Background: S1P1 (or EDG1) is a widely distributed G-protein-coupled receptor for sphingosine-1-phosphate (S1P), a blood-borne bioactive lipid. Stimulation of the S1P1 receptor triggers a Gi-linked pathway, leading to growth, survival, migration, and morphogenesis. Disruption of the S1P1 gene in mice results in embryonic lethality because of its key role within endothelial cells in regulating the coverage of blood vessels by vascular smooth muscle cells. S1P1 also mediates activation of Rac and functions as a typical chemotactic receptor.
Application: Functional assays
Figure 1. Dose-dependent inhibition of forskolin stimulated intracellular cAMP accumulation upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 2. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Takuwa et al. (2001) Subtype-specific, differential activities of the EDG family receptors for sphingosine-1-phosphate, a novel lysophospholipid mediator. Mol Cell Endocrinol 177:3-11.