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Description

Millipore's Ready-to-Assay GPCR frozen cells are designed for simple, rapid calcium assays with no requirement for intensive cell culturing. Millipore has optimized the freezing conditions to provide cells with high viability and functionality post-thaw. The user simply thaws the cells and resuspends them in media, dispenses cell suspension into assay plates and, following over night recovery, assays for calcium response. 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). Millipore's cloned human C3a-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant C3a expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists, antagonists, and modulators at C3a.