MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT GHRH RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human GHRHR cDNA (GenBank Accession Number NM_000823) with FLAG tag sequence at N-terminus.
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable after minimum of two months continuous growth
Background: Growth hormone-releasing hormone (GHRH), acting through the GHRH receptor GHRHR, plays a pivotal role in the regulation of GH synthesis and secretion in the pituitary. The isolated cDNA of the GHRHR predicted a 423-amino acid protein that has 7 putative transmembrane domains, characteristic of G protein- coupled receptors. It is a member of the secretin family of G protein-coupled receptors and has 47%, 42%, 35%, and 28% identity with receptors for vasoactive intestinal peptide, secretin, calcitonin and parathyroid hormone, respectively.
Application: Functional assays
Figure 1. Dose-dependent accumulation of intracellular cAMP upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). 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.
Gaylinn et al. (1993) Molecular cloning and expression of a human anterior pituitary receptor for growth hormone-releasing hormone. Molec. Endocr 7:77-84.
DeAlmeida and Mayo (1998) Identification and binding domains of the growth hormone-releasing hormone receptor by analysis of mutant and chimeric receptor proteins. Molec Endocr 12:750-765.