MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT LPA3 (EDG7) RECEPTOR
1 vial (1 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human LPAR3 cDNA (GenBank Accession Number NM_012152) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 3 μg/mL puromycin
Stable in culture for a minimum of 2 months
Background: LPA and the structurally related lysophoslipid mediator sphingosine 1- phosphate (S1P) signal cells through a set of G protein-coupled receptors known as EDG receptors. Some EDG receptors (e.g., EDG1) are S1P receptors; others (e.g., EDG2) are LPA receptors. LPA3 receptor (EDG7) mediates responses preferentially to unsaturated LPA, whereas LPA2 receptor (EDG4) mediates responses to both saturated and unsaturated LPA.
Application: Functional assays
Figure 1. Dose-dependent calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Figure 2. Dose-dependent inhibition of forskolin-stimulated intracellular cAMP level upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 3. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Bandoh et al. (1999) Molecular cloning and characterization of a novel human G-protein-coupled receptor, EDG7, for lysophosphatidic acid. J. Biol. Chem.274:27776-27785.
Ye et al. (2005) LPA3-mediated lysophosphatidic acid signalling in embryo implantation and spacing. Nature 435:104-108.