Application

Radioligand binding assay (See images & graphs link)

Biochem/physiol Actions

Target Sub-Family: Anaphylotoxin

GPCR Class: A

Protein Target: C5aR

Disclaimer

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.

General description

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 & 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.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

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).

Quality

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

Storage and Stability

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