MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT GPR35 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human GPR35 cDNA (GenBank Accession Number NM_005301) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM/F12, 10% FBS, 1 µg/mL puromycin, 1 mg/mL G418
Stable after minimum of two months continuous growth
Background: GPR35 is an orphan receptor and expressed in various tissues including stomach, gastrointestinal tissues, and several types of immune cells. Up- regulation of GPR35 has been found in human mast cells upon stimulation with IgE antibodies, human macrophages treated with the environmental contaminant polycyclic aromatic hydrocarbon benzo[a]pyrene, failing heart cells, and gastric cancer cells. Known agonists of the orphan receptor GPR35 are kynurenic acid, zaprinast, 5-nitro-2-(3-phenylproplyamino) benzoic acid, and lysophosphatidic acids.
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.
Wang JH et al. (2006) Kynurenic Acid as a Ligand for Orphan G Protein-coupled Receptor GPR35. J Biol Chem 281:22021-22028.
MacKenzie1, AE, et al. (2011) GPR35 as a novel therapeutic target. Frontiers in Endocrinology, 2:1-10.
Zhao PW, et al. (2010) Targeting of the Orphan Receptor GPR35 by Pamoic Acid: A Potent Activator of Extracellular Signal-Regulated Kinase and beta-Arrestin2 with Antinociceptive Activity. Mol Pharm 78:560–568.