MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT P2Y13 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human P2Y13 cDNA (GenBank Accession Number NM_023914) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable after two month continuous growth
Background: GPR86 or P2Y13 is a 333-amino acid 7 transmembrane protein that lacks a leader peptide but posseses a DRF motif. Northern blot and PCR analysis has shown GPR86 to be expressed highly in the spleen, with weaker expression in placenta, leukocytes, brain, placenta, lung, liver, spinal cord, thymus, small intestine, uterus, stomach, testis, fetal brain and adrenal glad and no expression in the pancreas, heart, kidney, skeletal muscle, ovary or fetal aorta. Communi et al determined that GPR86 shows a high affinity for ADP through pharmacologic charactization of GPR86-transfected human astrocytoma cells and CHO cells.
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). 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.
Communi et al. (2001) Identification of a novel human ADP receptor coupled to G(i). J Biol Chem 276:41479-41485
Lee et al. (2001) Discovery and mapping of ten novel G proteincoupled receptor genes. Gene 275:83-91.
Wittenberger et al. (2001) An expressed sequence tag (EST) data mining strategy succeeding in the discovery of new G-protein coupled receptors. J Molec Biol307:799-813.