ChemiSCREEN™ CCR9 Calcium-Optimized FLIPR Cell Lines
Description:
ChemiSCREEN™ CCR9 Calcium-Optimized FLIPR Cell Lines
Trade Name:
ChemiScreen
Product Overview:
TRANSFECTION: Full-length human CCR9 cDNA
Background Information:
The CCR9 receptor is a GPCR expressed selectively in the thymus, lymph node, spleen, and the small bowel (SB). CCR9 binds to Thymus-expressed chemokine ligand (TECK / CCL25) (Zaballos et al. 1999, Yu et al., 2000, Carramolino et al., 2001), and is involved in directing the movement of mature T cells in the thymus. CCR9 also plays an important role in the pathogenesis of SB Crohn's disease (Papadakis et al, 2001). The CCR9 selective expression in the small bowel but not the colon indicates the the role of CCR9 in the specialized immune response in the SB. It was also shown that CCR9-mediated antiapoptosis could help in the study of cell survival mechanism and T cell malfunctioning. Chemicon's cloned human CCR9-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CCR9 expression on the cell surface and contains high levels of the promiscuous Galpha15 protein to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between CCR9 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 10e6 cells/mL
Host Cells:
Chem-1, an adherent cell line expressing the promiscuous G protein, Gα15.
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:
- CCR9
- GPR28
- CC-CKR-9
- D6
- GPR-9-6
- CDw199
- CCR10
- CCR-9
- CMKBR9
- CCBP2
Format:
Cell Line
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 TECK: ~ 7.7 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.