ChemiSCREEN C5aR Membrane Preparation, recombinant

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 & 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 membrane preparations are ideal tools for screening for antagonists of C5a/C5aR interactions. The C5aR Membrane Preps bind specifically to 125I-labeled ligand C5a, with Kd of 0.59 nM. With 5 μg membrane per well and 0.1 nM 125I-labeled C5a, more than 15-fold signal:background was obtained.

Full-length human C5aR cDNA

Radioligand binding assay (See images & graphs link)

GPCR Class: A

Protein Target: C5aR

Target Sub-Family: Anaphylotoxin

Membranes were adjusted to 0.5 mg protein/ml in 50 mM Tris pH 7.4, 10% glycerol and 1% BSA, rapidly frozen, and stored at -80°C. One vial contains 1000 units, where a unit is the amount of membrane that will yield greater than 1000 cpm specific binding of I-125 labeled C5a (0.1 nM input concentration).

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

SPECIFICATIONS: Bmax for [¹²⁵I]C5a binding: 9.7 pmol/mg protein
Kd for [125I] C5a binding: ~ 0.59 nM

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