MULTISCREEN™ STABLE CELL LINES
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Full-length Human GHSR cDNA (GenBank Accession Number NM_198407.1) with FLAG-tag sequence at the N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable in culture for minimum of two months
Background: The ghrelin receptor is the target of growth hormone secretagogues, a class of synthetic peptide and non-peptide compounds that stimulate growth hormone (GH) release from the anterior pituitary. Ghrelin, the endogenous ligand for the ghrelin receptor, is predominantly secreted from X/A-like cells within the gastric mucosaandmaybethesourceofthemajorityofcirculatingplasmaghrelin. Ghrelin stimulates gastric acid secretion and motility, and may have significant effects on appetite and energy. It is not only important for the acute regulation of food intake but also plays an important role in the regulation of long term energy homoeostasis. Ghrelin has a number of actions in cardiovascular system, consistent with the localization of receptors to cardiovascular tissue.
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 accumulation of intracellular IP1 upon treatment with ligand, measured with IP-one Tb kit (Cisbio 62IPAPEC). 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.
Howard et al. (1996) A receptor in pituitary and hypothalamus that functions in growth hormone release. Science 273:974-977.
Kojima and Kangawa (2005) Ghrelin: structure and function. Physiol Rev 85:495-522.
van der Lely et al. (2004) Biological, physiological, pathophysiological, and pharmacological aspects of ghrelin. Endocr Rev 25:426-457.