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Calcium flux in OX1-expressing Chem-1 cell line induced by Orexin A. OX1-expressing Chem-1 cells and Wild-Type Chem-1 cells (Ch ...
ChemiScreen™ Human Recombinant OX1 Orexin Receptor Calcium-Optimized Stable Cell Line
Description:
ChemiScreen™ Human Recombinant OX1 Orexin Receptor Calcium-Optimized Stable Cell Line
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Product Overview:
Full-length human HCRTR1 cDNA encoding OX1.
Background Information:
The orexin/hypocretin peptides orexin A and orexin B (also known as hcrt-1 and hcrt-2) are 33- and 28-amino acid peptides, respectively, that are preferentially expressed in hypothalamus. The orexins bind to two GPCRs, OX1 and OX2, to increase intracellular calcium. Experiments with OX1 selective antagonists have shown the receptor plays a role in food consumption and also in acquisition and consolidation of spatial memory.(Haynes et al., 2000; Akbari et al, 2006) In addition, the orexinergic system plays a role in energy metabolism and stress responses (Kukkonen et al., 2002). Chemicon's cloned human OX1-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant OX1 expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between the OX1 and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Cell Line Type:
GPCR Cell Lines
Host Cells:
Chem-1, an adherent cell line expressing the promiscuous G protein, Gα15.
Packaging:
2 x 106 cells/vial
Materials Required but Not Delivered:
DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin.
Cell Type:
Chem-1
Gene Symbol:
- HCRTR1
- Ox1r
- OX1R
Presentation:
Cells are frozen at 2 x 106 cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by Orexin A: ~ 1 nM
Protein or Enzyme Type:
GPCRs
Species:
Human
Storage Conditions:
Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2.. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca++ and Mg++-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.