ChemiSCREEN™ C3aR Calcium-Optimized FLIPR Cell Line
Description:
ChemiSCREEN™ C3aR Calcium-Optimized FLIPR Cell Line
Trade Name:
ChemiScreen
Product Overview:
Full-length human C3aR cDNA
Background Information:
C3a, along with C4a and C5a, is a 77 amino acid anaphylotoxin generated by proteolytic cleavage during activation of the complement pathway (Ember and Hugli, 1997). The anaphylotoxins strongly promote inflammation by recruiting leukocytes, particularly basophils, eosinophils, neutrophils and monocyte (Martin et al., 1997). The proinflammatory properties of C3a are mediated by interaction between the peptide and a 7-TM G-protein coupled receptor, C3aR (Crass et al., 1996). Genetic and pharmacological inhibition of C3a/C3aR interaction indicates an important role for C3aR in allergic asthma (Humbles et al., 2000; Drouin et al., 2002). C3a/C3aR also enhances the effect of SDF-1 in promoting retention of haematopoietic stem/progenitor cells within the bone marrow (Rataiczak et al., 2004). Chemicon's cloned human C3aR-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant C3aR 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 C3aR and its ligand C3a.
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
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:
- C3AR1
- C3a-R
- HNFAG09
- C3AR
- C3R1
- AZ3B
Format:
Cell Line
Presentation:
Cells are frozen at 2 x 10E6 cells/mL in DMEM/20% fetal bovine serum containing 10% DMSO. Cell line tests negative for mycoplasma.
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by C3a: ~ 8 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 growth media, and replace media after 8-24 h. Cells are typically passaged 1:10 every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.