Application

Radioligand binding assay.

Biochem/physiol Actions

GPCR Class: A

Target Sub-Family: Chemokine

Protein Target: CXCR2 / IL-8b

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

TRANSFECTION: Full-length human CXCR2 cDNA (Accession Number: M73969)

CXCR2 is a 7-TM G-protein coupled receptor that binds to the chemokines GROalpha, GRObeta, GROgamma, IL-8, ENA-78, NAP-2 and GCP-2 (Olson and Ley, 2002). Neutrophils, mast cells and microvascular endothelial cells express CXCR2, and interactions of CXCR2 with its ligands promotes chemotaxis of these cell types (Heidemann et al., 2003; Nilsson et al., 1999; White et al., 1998). Studies with mice lacking CXCR2 indicate that CXCR2 promotes growth of primary tumors and secondary metastases (Keane et al., 2004), and plays an essential role in hyperoxia-induced lung injury (Sue et al., 2004). In addition, cytomegalovirus encodes a CXCR2-binding chemokine, vCXC-1, that promotes neutrophil migration to infected cells (Penfold et al., 1999). Chemicon′s CXCR2 Membrane Preps are ideal tools for screening for antagonists of interactions between CXCR2 and its ligands. Radiolabeled IL-8 binds to the CXCR2 membranes with a Kd of 0.075 nM. With 0.1 nM 125I-labeled IL-8, a 5-fold window of specific binding is 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 typically contains 1000 units, where a unit is the amount of membrane that will yield greater than 1000 cpm specific binding of 125I-labeled IL-8 (0.1 nM input concentration).

Quality

Bmax for [¹²⁵I]IL-8 binding: 0.93 pmol/mg protein; Kd for [¹²⁵I] IL-8 binding: ~ 0.075 nM

Storage and Stability

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