MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT EP3 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human PTGER3 cDNA (GenBank Accession Number NM_000957) 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: Prostaglandin E2 (PGE2) is involved in a number of physiologic and pathophysiologic events in many tissues of the body. The biologic effects of PGE2 are mediated through interaction with specific membrane-bound G protein-coupled prostanoid EP receptors. EP3 receptor (or PTGER3) is expressed as multiple transcripts through alternative splicing, with each transcript showing a different tissue-specific distribution. PGE2 may mediate fever generation in response to both endogenous and exogenous pyrogens by acting at the EP3 receptor. EP3-mediated neuronal pathways converge at corticotropin-releasing hormone containing neurons in the paraventricular nucleus of the hypothalamus to induce HPA axis activation during sickness.
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.
Adam et al. (1994) Cloning and expression of three isoforms of the human EP(3) prostanoid receptor. FEBS Lett 338:170-174.
Matsuoka et al. (2003) Impaired adrenocorticotropic hormone response to bacterial endotoxin in mice deficient in prostaglandin E receptor EP1 and EP3 subtypes. Proc Nat Acad Sci USA 100:4132-4137.
Ushikubi et al. (1998) Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP(3). Nature 395:281-284.