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HTS054M ChemiScreen™ CXCR6 Membrane Preparation

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HTS054M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-5CXCR6
      Description
      Catalogue NumberHTS054M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ CXCR6 Membrane Preparation
      OverviewHuman CXCR6 cDNA
      Background InformationThe GPCR CXCR6 (previously known as BONZO, STRL33 and TYMSTR) binds selectively to the free chemokine domain of CXCL16, which is derived from a membrane-bound precursor containing a CXC-containing chemokine domain, a glycosylated mucin-like domain, and a transmembrane domain (Wilbanks et al., 2001) CXCR6 is selectively expressed on Th1, Th2 and Tr1 T cell subsets, whereas CXCL16 is expressed on monocytes/macrophages and dendritic cells (Tabata et al., 2005). CXCR6 promotes migration of activated lymphocytes to sites of inflammation in tissues such as liver and synovium (Nanki et al., 2005, Sato et al., 2005). Chemicon's CXCR6 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 CXCR6 interactions with its ligands. The membrane preparations exhibit a Kd of 0.6 nM for [125I]-CXCL16. With 2.5 or 10 μg/well CXCR6 Membrane Prep and 0.5 nM [125I]-CXCL16, greater than 4-fold and 9-fold signal-to-background ratios are obtained, respectively.
      References
      Product Information
      FormatMembranes
      PresentationOne vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific CXCL16-stimulated [35S]-GTPγS binding. If radioligand binding assays are desired, it is recommended to double the mass of membrane prep per well to increase signal window..

      Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.

      Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
      Quality LevelMQ100
      Applications
      ApplicationHuman CXCR6 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay, and GTPγS binding.
      Biological Information
      Entrez Gene Number
      Gene Symbol
      • CXCR6
      • CDw186
      • BONZO
      • CXC-R6
      • CXCR-6
      • TYMSTR
      • CD186
      • STRL33
      GPCR ClassA
      Host CellsChem-5
      Protein TargetCXCR6
      Target Sub-FamilyChemokine
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: O00574 # Receptor for the C-X-C chemokine CXCL16. Used as a coreceptor by SIVs and by strains of HIV-2 and m-tropic HIV-1.
      SIZE: 342 amino acids; 39280 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in lymphoid tissues and activated T cells.
      SIMILARITY: SwissProt: O00574 ## 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, a GF/C 96-well filter plate 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] CXCL16 (Perkin Elmer catalog # NEX398)

      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 4-fold signal:background with 125I-labeled CXCL16 at 0.5 nM
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceSPECIFICATIONS:
      1 unit = 5 μg for GTPγS binding assay
      EC50 in GTPγS binding assay by CXCL16: 0.52 – 1.2 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 ConditionsStore at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      HTS054M 04053252677854

      Documentation

      ChemiScreen™ CXCR6 Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Distribution and kinetics of SR-PSOX/CXCL16 and CXCR6 expression on human dendritic cell subsets and CD4+ T cells.
      Tabata S et al.
      J. Leukoc. Biol. , 77:777-86 (2005)  2005

      15703197 15703197
      Pathogenic role of the CXCL16-CXCR6 pathway in rheumatoid arthritis.
      Nanki, Toshihiro, et al.
      Arthritis Rheum., 52: 3004-14 (2005)  2005

      Show Abstract
      16200580 16200580
      Role for CXCR6 in recruitment of activated CD8+ lymphocytes to inflamed liver.
      Sato T et al.
      J. Immunol., 174:277-83 (2005)  2005

      15611250 15611250
      Expression cloning of the STRL33/BONZO/TYMSTR ligand reveals elements of CC, CXC, and CX3C chemokines.
      Wilbanks A et al.
      J. Immunol., 166:5145-5154 (2001)  2001

      11290797 11290797

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      Categories

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