ChemiScreen™ ChemR23 Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ ChemR23 Calcium-Optimized FLIPR Cell Lines
Trade Name:
ChemiScreen
Product Overview:
Full-length human ChemR23 cDNA
Background Information:
ChemR23 was discovered as an orphan receptor related to the chemoattractant receptors C3a, C5a and FPR1, and expressed on dendritic cells and macrophages (Samson et al., 1998). A ligand for ChemR23 was characterized as chemerin, a 15 kD proteolytically processed protein found in inflammatory sites; a 9 amino acid peptide from the C-terminus of chemerin is sufficient to activate ChemR23 (Wittamer et al., 2003, 2004). Chemerin expressed in lymphoid and microvascular endothelium mediates migration of ChemR23-expressing dendritic cells to lymphoid organs and vasculature at sites of inflammation (Vermi et al., 2005). In addition, a bioactive lipid, resolvin E1, was found to functionally interact with ChemR23 to reduce inflammation (Arita et al., 2005). Chemicon's cloned human ChemR23-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant ChemR23 expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between ChemR23 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
Concentration:
2 x 106 cells/mL
Host Cells:
Chem-1, an adherent cell line expressing the promiscuous G protein, Gα15.
Packaging:
1 mL per vial
Materials Required but Not Delivered:
GROWTH MEDIA: 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:
- CMKLR1
- ChemR23
- DEZ
- MGC126106
- MGC126105
- CHEMR23
Presentation:
Cells are frozen at 2 x 10e6 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 Chemerin: ~ 4.7 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.