ChemiSCREEN CX3CR1 Membrane Preparation

General description

CX3CR1 is a GPCR expressed on natural killer cells, cytotoxic T lymphocytes, and macrophages. The sole ligand for CX3CR1, fractalkine, is an unusual chemokine that is expressed as a transmembrane molecule with a CX3C domain and a mucin domain (Imai et al., 1997). Fractalkine is highly expressed on endothelial cells activated by TNFa and other proinflammatory cytokines, and fractalkine/CX3CR1 interactions mediate recruitment of macrophages into the atherosclerotic plaque (Lesnick et al., 2003; McDermott et al., 2003). In addition, fractalkine and CX3CR1 have been implicated in the pathogenesis of glomerulonephritis, HIV infection and rheumatoid arthritis (Ito et al., 2002; Faure et al., 2003; Nanki et al., 2002). Chemicon′s CX3CR1 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of fractalkine/CX3CR1 interactions. The membrane preparations exhibit a Kd of 0.05 nM for [¹²⁵I]-fractalkine. With 2.5 mg/well CX3CR1 Membrane Prep and 0.05 nM [¹²⁵I]-fractalkine, a greater than 7-fold signal-to-background ratio is obtained.

Full-length Human CX3CR1

Application

Human CX3CR1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

Radioligand binding assay, and GTPγS binding.

Biochem/physiol Actions

GPCR Class: A

Protein Target: CX3CR1

Target Sub-Family: Chemokine

Quality

Bmax: 3.13 pmol/mg

Kd: 0.05 nM

Specifications

Inucbation Conditions
RECOMMENDED ASSAY CONDITIONS:

Binding buffer: 50mM HEPES, 5mM MgCl₂, 1mM CaCl₂, 0.2% BSA, pH to 7.4 and filter.

Radioligand: [¹²⁵I] Fractalkine (PerkinElmer#: NEX368)

One vial contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 5-fold signal:background with ¹²⁵I-labeled fractalkine at 0.05 nM.

Physical form

Packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA

Packaging method: Membranes protein were adjusted to 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.

Storage and Stability

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

Legal Information

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

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.