ChemiScreen CXCR2 Membrane Preparation

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

Radioligand binding assay.

GPCR Class: A

Protein Target: CXCR2 / IL-8b

Target Sub-Family: Chemokine

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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.

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

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).

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

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