GeneBLAzer® HCRTR2-Gα15 CHO-K1 DA Assay Kit

Catalog number: K1369

 Related applications: Cellular Imaging | Target & Lead Identification & Validation

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Description

This cell line is available as dividing cells by requesting a quote for 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 (#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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

System: GeneBLAzer®
Gene ID: 3062
Reporter: BLA (Beta-Lactamase)
Cell Line: CHO-K1
Cell State: Division-Arrested Cells
Gene Symbol: HCRTR2
Product Size: 1 plate
Accession Number: NM_001526.2
Detection Method: Fluorescent
Druggable Target: GPCRs
Population Status: Single Cell Clone Origin

Contents & storage




Storage:

GeneBLAzer® HCRTR2-Gα15 CHO-K1 DA Cells and HCRTR2-Gα15-NFAT-bla CHO-K1 Cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt or thaw for immediate use.


Contents:
GeneBLAzer® HCRTR2-Gα15 CHO-K1 DA Assay Kit
Each system contains sufficient division arrested (DA) cells & substrate to assay 1 x 384-well plate. (Other materials are required separately; please refer to the protocol).
Includes:
•HCRTR2-Gα15 CHO-K1 DA cells
•LiveBLAzer™-FRET B/G Loading Kit, 70µg
•Solution D, 1ml
•2 protocols
•Certificate of Analysis
GeneBLAzer® HCRTR2-Gα15 CHO-K1 DA cells
Each system contains sufficient division arrested (DA) cells to assay 10 x 384-well plates (LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•HCRTR2-Gα15 CHO-K1 DA cells
•1 protocol
•Certificate of Analysis

GeneBLAzer® HCRTR2-Gα15-NFAT-bla CHO-K1 cells
(LiveBLAzer™-FRET B/G Loading Kit, Solution D and other materials are required separately; please refer to the protocol).
Includes:
•HCRTR2-Gα15-NFAT-bla CHO-K1 cells
•1 protocol
•Certificate of Analysis

Documents

Manuals & protocols