Thermo Scientific™ GeneBLAzer™ HCRTR2-Gα15 CHO-K1 DA Assay Kit

Description

This cell line is available as dividing cells by requesting a quote for Cat. No. K1246 from dividingcells@invitrogen.com

GeneBLAzer™ HCRTR2-Gα15 CHO-K1 DA(Division Arrested) cells and HCRTR2-Gα15-NFAT-bla CHO-K1 cells contain the human Hypocretin 2 receptor (HCRTR2) stably integrated into the GeneBLAzer™ Ga15-NFAT-bla CHO-K1 cell line. Gα15-NFAT-bla CHO-K1 cells (Cat. No. K1213) contain a beta-lactamase reporter gene under control of the nuclear factor of activated T-cells Response Element (NFAT), as well as the promiscuous G-protein, Ga15. Division Arrested (DA) cells are available in two configurations: an Assay Kit (which includes cells and sufficient substrate to analyze 1 x 384-well plate), and a tube of cells sufficient to analyze 10 x 384-well plates.

DA cells are irreversibly division arrested using a low-dose treatment of Mitomycin-C, and have no apparent toxicity or change in cellular signal transduction. Both HCRTR2-Gα15 CHO-K1 DA cells and HCRTR2-Gα15-NFAT-bla CHO-K1 cells are functionally validated for Z'-factor and EC₅₀ concentrations of Orexin B (Figure 1). In addition, HCRTR2-Gα15-NFAT-bla CHO-K1 cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time (data available upon request). Additional testing data using alternate stimuli are also available.

HCRTR2 has high affinity for both orexin peptides. Expression of this receptor has been detected in the brain as well as adrenal gland, gastrointestinal tract, and pancreas. The signaling cascades used by orexin receptors appear to be varied and cell type dependant. In CHO cells, calcium release is seen after agonist treatment. In hypothalamic neurons, the frequency of action potentials is increased. Administering either orexin peptide into the bloodstream of rats caused an increase in feeding behavior. The orexin system is also thought to regulate neuroendocrine function and the sleep wake cycle. HCRTR2 has been directly implicated in narcolepsy in animal studies.