MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT MGLUR7 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human GRM7 cDNA (GenBank Accession Number NM_000844) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM with GlutaMAX (Gibco 10566), 10% FBS (dialyzed), 2 mM sodium pyruvate, 1 μg/mL puromycin
Stable after two months of continuous growth
Background: The metabotropic glutamate receptors (mGluRs) are divided into 3 groups on the basis of sequence homology, putative signal transduction mechanisms, and pharmacological properties. Group II and group III mGluRs are linked to inhibition of the cyclic AMP cascade, but differ in their agonist selectivity. mGluR7 is a member of Group III. Immunohistochemistry and in situ hybridization showed that mGluR7 expresses in various sites in the nervous system.
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. Dose- dependent response 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 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.
Wu et al. (1998) Group III human metabotropic glutamate receptors 4, 7, and 8: molecular cloning, functional expression, and comparison of pharmacological properties in RGT cells. Brain Res Mol Brain Res 53:88-97.
Makoff et al. (1996) Human metabotropic glutamate receptor type: molecular cloning and mRNA distribution in the CNS. Brain Res Mol Brain Res 40:165-170.
Muto et al.(2007) Structures of the extracellular regions of the group II/III metabotropic glutamate receptors. Proc Natl Acad Sci USA 104:3759-3764.