MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT S1P5 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human S1P5 cDNA (GenBank Accession Number: NM_030760) 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: The lysosphingolipid sphingosine 1-phosphate (S1P) regulates cell proliferation, apoptosis, motility, and neurite retractions. S1P and the structurally related LPA signal cells through a set of G-protein-coupled receptors known as EDG receptors. S1P5 or EDG8 is a 400-amino acid 7 transmembrane protein that shares 42-49% amino acid identity with the human S1P receptors EDG1, EDG3, and EDG5. Northern blot analysis has shown widespread expression of EDG8 in rat tissues, with strongest expression in spleen and white matter tracts of the brain.
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.
IM et al. (2000) Characterization of a novel phingosine 1-phosphate receptor, Edg-8. J Biol Chem 275:14281-14286.