MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT GPR109B RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human GPR109B cDNA (GenBank Accession Number NM_006018.1) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
DMEM/F12, 10% FBS, 10 μg/mL puromycin
Stable after minimum two months of continuous growth
Background: GPR109B (or HM74) is a 387-amino acid, 7-transmembrane protein responsible for the directed migration of specific cell types. GPR109B has highest expression in spleen, lymphocytes and adipose tissue, and lower expression in the heart, placenta, prostate and bone marrow. Nicotinic acid has been used as a lipid-lowering agent possibly by acting through GPR109A and GPR109B in adipose tissues. This supports the suggestion that signaling through this nicotinic acid receptors may be a mechanism for the treatment of hyperlipidemia and insulin-resistant states.
Application: Functional assays
Figure 1. 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 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.
Karpe and Frayn (2004) The nicotinic acid receptor-a new mechanism for an old drug. Lancet 363:1892-1894.
Tunaru et al. (2003) PUMA-G and HM74 are receptors for nicotinic acid and mediate its anti- lipolytic effect. Nature Med 9:352-355.
Wise et al. (2003) Molecular identification of high and low affinity receptors for nicotinic acid. J Biol Chem 278:9869-9874.
Nomura et al. (1993) Molecular cloning of cDNAs encoding a LD78 receptor and putative leukocyte chemotactic peptide receptors. Int Immun 5:1239-1249.