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HTS003M ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR3 Chemokine Receptor

Human CXCR3 GPCR membrane preparation for Radioligand binding Assays.

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Host Cell: Chem-1 Uni Prot Number: P49682 Entrez Gene Number: NM_001504.1
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HTS003M
200 units  
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1CXCR3
      Description
      Catalogue NumberHTS003M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human CXCR3 Chemokine Receptor
      OverviewTRANSFECTION: Full-length human CXCR3 cDNA
      Background InformationCXCR3 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.
      References
      Product Information
      FormatMembranes
      PresentationLiquid 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.
      Quality LevelMQ100
      Applications
      ApplicationHuman CXCR3 GPCR membrane preparation for Radioligand binding Assays.
      Key Applications
      • Radioligand binding assay
      Application NotesRadioligand binding assay (See images & graphs link)
      Biological Information
      Concentration1.0 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryCD183 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.
      Gene Symbol
      • CXCR3
      • CD182
      • CXC-R3
      • IP10-R
      • IP10
      • Mig-R
      • CKR-L2
      • CD183
      • CMKAR3
      • GPR9
      • CXCR-3
      • MigR
      GPCR ClassA
      Host CellsChem-1
      Protein TargetCXCR3
      Target Sub-FamilyChemokine
      UniProt Number
      UniProt SummaryFUNCTION: 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.
      Incubation ConditionsRECOMMENDED 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
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with CXCR3 membrane prep.
      Product Usage Statements
      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 Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Bestellnummer GTIN
      HTS003M 04053252743504

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      Kategorien

      Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations