MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT GPR68 RECEPTOR
1 vial (2 x 106) frozen cells
Cellbanker 2 (Amsbio 11891)
Expression vector containing full-length human GPR68 cDNA (GenBank Accession Number NM_003485) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable for a minimum of 1 month in continuous culture
Background: GPR68, also known as ovarian cancer G-protein-coupled receptor 1 (OGR1), was previously considered a receptor for sphingosylphosphorylcholine. Recent studies revealed that GPR68 is a proton-sensing GPCR that plays a major role in pH homeostasis. The receptor is expressed in the kidney, placenta, heart, lung, brain, spleen, testis, small intestines, and peripheral blood leucocytes. GPR68 stimulates inositol phosphate (IP) production, Ca2+ mobilization, and cAMP accumulation. The receptor is involved in cell-mediated responses to acidosis in the bone, acts as a metastasis suppressor gene in prostate cancer, and is a potential therapeutic target for obstructive lung diseases.
Application: Functional assays
Figure 1. Dose-dependent calcium flux upon treatment with pH, with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Figure 2. Dose-dependent increase of intracellular cAMP level upon treatment with pH, 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.
Ludwig, M.-G., Vanek, M., Guerini, D., Gasser, J. A., Jones, C. E., Junker, U., Hofstetter, H., Wolf, R. M., Seuwen, K. Proton-sensing G-protein-coupled receptors. Nature 425: 93-98, 2003.
Saxena, H., Deshpande, D., Tiegs, B., Yan, H., Battafarano, R., Burrows, W., Penn, R. (2012). The GPCR OGR1 (GPR68) mediates diverse signalling and contraction of airway smooth muscle in response to small reductions in extracellular pH. British Journal of Pharmacology, 166(3), 981–990.
YangM, et al.(2006) Expression of and role for ovarian cancer G-protein–coupled receptor 1 (OGR1) during osteoclastogenesis. J Biol Chem 281(33):23598–23605.