ChemiSCREEN™ C5aR Calcium-Optimized FLIPR Cell Line

ChemiSCREEN™ C5aR Calcium-Optimized FLIPR Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human C5aR cDNA

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.

Calcium flux assay, ligand binding assays

Human

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.

FUNCTION: SwissProt: P21730 # Receptor for the chemotactic and inflammatory peptide anaphylatoxin C5a. This receptor stimulates chemotaxis, granule enzyme release and superoxide anion production.
SIZE: 350 amino acids; 39321 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
PTM: Sulfation plays a critical role in the association of the receptor with C5a, but no significant role in the ability of the receptor to transduce a signal and mobilize calcium in response to a small peptide agonist.
SIMILARITY: SwissProt: P21730 ## Belongs to the G-protein coupled receptor 1 family.

Human

Chemicon

C5aR

Cells are frozen at 2 x 106 cells/mL in growth media containing 10% DMSO. Cell line tests negative for mycoplasma.

peptide

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.

Anaphylotoxin

A

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• C5AR1
• CD88
• C5AR
• C5A
• C5R1
• C5aR
• C5a-R

GPCRs

ChemiSCREEN™ C5aR Calcium-Optimized FLIPR Cell Line

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

2 x 10(6) cells/mL

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

Chem-1

GPCR Cell Lines

Chem-1

Cell Line

P21730

2x10(e6) cells/vial

NM_001736.3