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Calcium flux in CCR4-expressing Chem-5 cell line induced by TARC. CCR4-expressing Chem-5 cells and wild-type Chem-5 cells were loaded with Fluo-4 and ...
ChemiScreen™ CCR4 Calcium-Optimized FLIPR Cell Line
Description:
ChemiScreen™ CCR4 Calcium-Optimized FLIPR Cell Line
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Product Overview:
Full-length human CCR4 cDNA
Background Information:
CCR4 is a GPCR that is activated by chemokines TARC (thymus and activation regulated chemokine) and MDC (macrophage-derived chemokine) (Olsen and Ley, 2001). NK cells, Th2 cells, cutaneous memory T cells, macrophages and platelets express CCR4 and respond to TARC and MDC (Andrew et al., 2001; Inngjerdingen et al., 2000; Gear et al., 2001; Soler et al., 2003). CCR4-null mice display reduced airway hyperresponsiveness to fungal spores, lipopolysaccharide-induced endotoxic shock, and cardiac allograft rejection (Chvatchko et al., 2000; Schuh et al., 2002; Huser et al., 2005). In addition, antibody -mediated antagonism of MDC inhibits development of insulitis and diabetes in NOD mice (Kim et al., 2002). Chemicon's cloned human CCR4-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CCR4 expression on the cell surface and contains high levels of the promiscuous G protein Ga15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between CCR4 and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays
Species Reactivity:
Human
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-5, an adherent cell line with high levels of the promiscuous G protein.
Packaging:
2 x 106 cells/mL
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-5
Gene Symbol:
- CCR4
- CMKBR4
- ChemR13
- K5-5
- k5-5
- CKR4
- HGCN:14099
- MGC88293
- CCR-4
- CC-CKR-4
Format:
Cell Line
Presentation:
Cells are frozen at 2 x 10(e6) cells/mL in DMEM/20% fetal bovine serum/100U/mL each penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.
Sales Restrictions:
This product requires a User Agreement (Label License). Please see product datasheet for further information
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by TARC: 0.59 nM
Signal/noise at TARC Emax: 524
Z' = 0.85 with TARC at 2x EC50
Signal/noise at TARC Emax: 524
Z' = 0.85 with TARC at 2x EC50
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 37°C 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 growth media, and replace media after 8-24 h. Cells are typically passaged 1:10 with 0.05% Trypsin/0.2 g/L EDTA every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.