ChemiSCREEN™ C5aR Calcium-Optimized FLIPR Cell Line
Description:
ChemiSCREEN™ C5aR Calcium-Optimized FLIPR Cell Line
Trade Name:
ChemiScreen
Product Overview:
Full-length human C5aR cDNA
Background Information:
C5a is a proinflammatory peptide generated through activation of the complement system, and is more potent than the other anaphylatoxins, C4a and C3a, in activating peripheral blood leukocytes (Gerard and Gerard, 1994). It can actively participate in the regulation of local cytokine network by stimulating production of proinflammatory cytokines (Buchner et al., 1995; Hsu et al., 1999). The proinflammatory effects of C5a are mediated through binding to a specific 7-TM chemoattractant receptor, C5aR (CD88). Binding of C5a to phagocyte C5aR induces chemotaxis, production of superoxide anions, and release of degradative enzymes. Pharmacologic or genetic disruption of C5a/C5aR interaction reduces sepsis (Riedemann et al., 2003), immune complex-induced lung disease (Shushakova et al., 2002), and Arthrogen-induced arthritis (Grant et al., 2002) in experimental models. Chemicon's cloned human C5a receptor-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant C5aR 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 C5a/C5aR interactions.
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 10(6) cells/mL
Host Cells:
Chem-1
Packaging:
2x10(e6) 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:
- C5AR1
- CD88
- C5AR
- C5A
- C5R1
- C5aR
- C5a-R
Format:
Cell Line
Presentation:
Cells are frozen at 2 x 106 cells/mL in growth media containing 10% DMSO. Cell line tests negative for mycoplasma.
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by C5a: ~ 0.6 nM
HOST CELLS:
TRANSFECTION: (Accession Number: M62505)
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
HOST CELLS:
TRANSFECTION: (Accession Number: M62505)
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
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 2 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 growth media. Cells are typically split 1:10 every 3-4 days.