MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT C5A RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human C5aR1 cDNA (GenBank Accession Number NM_001736) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable for a minimum of 2 months in continuous culture
Background: C5a receptor (C5a anaphylatoxin chemotactic receptor, or CD88 antigen, C5R1) is a receptor for chemotactic and inflammatory peptide anaphylatoxin C5a. C5a is a classic, potent inflammatory mediator, and the C5R1 receptor stimulates chemotaxis, granule enzyme release and superoxide anion production. C5a receptor antagonists, such as AcPhe[Orn-Pro-D-cyclohexylalanine-Trp-Arg], show therapeutic effects in various inflammatory disease models. C5a receptor may also be involved in neurodegeneration and Alzheimer’s disease.
Application: Functional assays
Figure 1. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Figure 2. Dose-dependent inhibition of forskolin-stimulated intracellular cAMP accumulation upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 3. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Sumichika, H. (2004). “C5a receptor antagonists for the treatment of inflammation.” Curr Opin Investig Drugs 5(5): 505-10.Woodruff, T. M., T. V. Arumugam, et al. (2003). “A potent human C5a receptor antagonist protects against disease pathology in a rat model of inflammatory bowel disease.” J Immunol 171(10): 5514-20.
Farkas, I., M. Takahashi, et al. (2003). “Complement C5a receptor-mediated signaling may be involved in neurodegeneration in Alzheimer’s disease.” J Immunol 170(11): 5764-71.