MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT DP2 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human DP2 cDNA (GenBank Accession Number NM_004778) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable after minimum of 2 months continuous growth
Background: The chemoattractant-homologous receptor expressed on TH2 cells (CRTH2, also known as DP2 and GPR44) serves as a receptor for prostaglandin D2 (PGD2) and has been reported to mediate PGD2-dependent migration of eosinophils. DP2 mediates the pro-inflammatory effects of PGD2 generated in allergic inflammation. 11-Dehydro- thromboxane B2 (11-dehydro-TXB2) was found to be a full agonist of the DP2 receptor and hence might cause the receptor activation in cellular contexts where PGD-synthase is not present. Antagonism of the 11-dehydro-TXB2/DP2 axis may be of therapeutic relevance.
Application: Functional assays
Figure 1. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Cells were transiently transfection with Gα16. Figure 2. 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 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.
Nagata et al. (1999) CRTH2, an orphan receptor of T-helper-2-cells, is expressed on basophils and eosinophils and responds to mast cell-derived factor(s). FEBS Lett 459:195- 199.
Bohm et al. (2004) 11-Dehydro-thromboxane B2, a stable thromboxane metabolite, is a full agonist of chemoattractant receptor-homologous molecule expressed on TH2 cells (CRTH2) in human eosinophils and basophils. J Biol Chem 279:7663-7670.