ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR3 Chemokine Receptor

ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR3 Chemokine Receptor

Chemicon (Millipore)

200 units

TRANSFECTION: Full-length human CXCR3 cDNA

CXCR3 is a 7-TM GPCR that is selective for the CXC chemokines IP10, ITAC and MIG (Loetscher et al., 1996). Binding of IP10 and MIG to CXCR3 induces Ca²⁺ mobilization, chemotaxis and inflammatory responses of T lymphocytes, and also act as potent inhibitors of angiogenesis. CXCR3 is highly expressed in IL-2-activated T lymphocytes in vitro (Loetscher et al., 1996), and in T lymphocytes present in inflamed tissues in rheumatoid arthritis and multiple sclerosis (Balashov et al., 1999; Qin et al., 1998). In vivo, neutralization of CXCR3 inhibits experimentally induced type I diabetes (Frigerio et al., 2002), peritonitis (Xie et al., 2003), and post-lung transplantation bronchiolitis obliterans syndrome (Belperio et al., 2002). Millipore’s ChemiScreenTM CXCR3 receptor membrane preparations are ideal tools for screening for antagonists of CXCR3 interactions with its ligands. With 0.1 nM ¹²⁵I-labeled IP10, the CXCR3 membrane preparation at 5 μg/well gives a signal:bakground of greater than 10-fold.

Radioligand binding assay (See images & graphs link)

Human

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.

CD183 is a G protein-coupled receptor with selectivity for three chemokines, termed IP10 (interferon-g-inducible 10 kDa protein), Mig (monokine induced by interferon-g) and I-TAC (interferon-inducible T cell a-chemoattractant). IP10, Mig and I-TAC belong to the structural subfamily of CXC chemokines, in which a single amino acid residue separates the first two of four highly conserved Cys residues. Historically, CD183 is the third CXC chemokine receptor discovered and, therefore, commonly designated as CXCR3. Binding of chemokines to CD183 induces cellular responses that are involved in leukocyte traffic, most notably integrin activation, cytoskeletal changes and chemotactic migration. Inhibition by Bordetella pertussis toxin suggests that heterotrimeric G protein of the Gi-subclass couple to CD183. Signal transduction has not been further analyzed but may include the same enzymes that were identified in the signaling cascade induced by other chemokine receptors. As a consequence of chemokine-induced cellular desensitization (phosphorylation-dependent receptor internalization), cellular responses are typically rapid and short in duration. Cellular responsiveness is restored after dephosphorylation of intracellular receptors and subsequent recycling to the cell surface. A hallmark of CD183 is its prominent expression in in vitro cultured effector/memory T cells, and in T cells present in many types of inflamed tissues. In addition, IP10, Mig and I-TAC are commonly produced by local cells in inflammatory lesion, suggesting that CD183 and its chemokines participate in the recruitment of inflammatory cells. Therefore, CD183 is a target for the development of small molecular weight antagonists, which may be used in the treatment of diverse inflammatory diseases.

FUNCTION: SwissProt: P49682 # Receptor for CXCL9, CXCL10 and CXCL11 and mediates the proliferation of human mesangial cells (HMC). Isoform 2 is a receptor for CXCL4 and also mediates the inhibitory activities of CXCL9, CXCL10 and CXCL11 on the growth of human microvascular endothelial cells (HMVEC). Isoform 2 may play a role in angiogenesis. Isoform 3 mediates activity of CXCL11.
SIZE: 368 amino acids; 40660 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Isoform 1 and isoform 2 are mainly expressed in heart, kidney, liver and skeletal muscle. Isoform 1 is also expressed in placenta.
SIMILARITY: SwissProt: P49682 ## Belongs to the G-protein coupled receptor 1 family.

Human

Chemicon

CXCR3

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.

peptide

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

Chemokine

A

For special offers Click here http://www.millipore.com/drugdiscovery/dd3/gpcrtargetsolutions

• CXCR3
• CD182
• CXC-R3
• IP10-R
• IP10
• Mig-R
• CKR-L2
• CD183
• CMKAR3
• GPR9
• CXCR-3
• MigR

GPCR Membrane Preparations

ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR3 Chemokine Receptor

1.0 mg/mL

Table 1. Signal:background and specific binding values obtained in a competition binding assay with CXCR3 membrane prep.

	10 µg/well
Signal:Background	28.2
Specific Binding (cpm)	6028

SPECIFICATIONS: 1 unit = 5 µg
Bmax for [¹²⁵I]IP-10 binding: 0.6 pmol/mg protein
Kd for [¹²⁵I]IP-10 binding: ~ 0.2 nM

Chem-1

RECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl₂, 1 mM CaCl₂, 0.2% BSA, filtered and stored at 4°C

Radioligand: [¹²⁵I] IP10 (Perkin Elmer #NEX348)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 10-fold signal:background with ¹²⁵I-labeled IP10 at 0.1 nM

Chem-1

Membranes

P49682

NM_001504.1

GPCR