MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT ETB RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Full-length Human EDNRB cDNA (GenBank Accession Number NM_000115) with FLAG-tag sequence at the N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DME/F12, 10% FBS, 10 µg/mL puromycin
Background: Endothelin is a hormone produced predominantly by endothelial cells that have been recognized to play a significant role in the development of several cardiovascular disease states. ETB is mainly expressed in vascular endothelial and and epithelial tissues as well as by smooth muscle cells. ETB activation of smooth muscle cells results in vasoconstriction, whereas ETB activation on vascular endothelium causes vasodilation through the release of nitric oxide. ETB has also been found to lower blood pressure through natriuresis and diuresis, and to release prostaglandins. In the kidney, ETB also serves to clear endothelins from circulation by receptor-mediated endocytosis and subsequent lysosomal degradation. Mutations in the ETB gene have been linked to Waardenburg syndrome, as well as Hirschsprung disease type 2.
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.
Frommer, KW et al. (2008) Expression and function of ETA and ETB receptors in SSc. Rheumatology 47 (Suppl 5): v27-v28.
Sato-Jin, K et al. (2008) Epistatic connections between microphthalmia-associated transcription factor and endothelin signaling in Waardenburg syndrome and other pigmentary disorders. FASEB J. 22(4):1155-68.
Tanaka, H et al. (1998) Novel mutations of the endothelin B receptor gene in patients with Hirschsprung’s disease and their characterization. J. Biol. Chem.273(18):11378-83.