MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT M4 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human CHRM4 cDNA (GenBank accession number NM_000741.2) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
DMEM/F12, 10% FBS, 10 μg/mL puromycin
Stable in culture for minimum of two months
Background: The human muscarinic M4 receptor is a 479-amino acid, 7- transmembrane protein. In heart muscle, M4 receptors are coupled to specific K+ currents that regulate cardiac muscle contractions. They can also be found in tissues of the central nervous system and bladder.
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 stimulation of intracellular cAMP accumulation 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.
Eglen (2006) Muscarinic receptor subtypes in neuronal and non-neuronal cholinergic function. Auton Atacoid Pharmcol 26:219-233.
Yeh et al. (2007) Atrial tachycardia induces remodeling of muscarinic receptors and their coupled potassium currents in canine left atrial and pulmonary vein cardiomyocytes. Br J Pharmacol 152:981-982.