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, 250ug/ml hygromycin
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 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.