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HTS001M | ChemiSCREEN™ Membrane Preparation Recombinant Human CXCR1 Chemokine Receptor

HTS001M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1 CXCR1 / IL-8a
      Description
      Catalogue NumberHTS001M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human CXCR1 Chemokine Receptor
      Overview: Full-length human CXCR1 cDNA
      Background InformationCXCR1 (also known as IL-8RA) is a Gi/o-coupled GPCR expressed on neutrophils, CD8(+) T cells, and intestinal epithelial cells (Murphy, 1997; Sturm et al., 2005; Takata et al., 2004). CXCR1 binds specifically to the chemokine interleukin-8 (IL-8), whereas a related receptor, CXCR2, binds promiscuously to IL-8 and several other chemokines. CXCR1 has been proposed to be the primary regulator of neutrophil response to IL-8 in sepsis (Cummings et al., 1999). Antagonists of CXCR1 and CXCR2 have been developed and display efficacy in animal models of reperfusion injury (Bertini, et al., 2004; Kaneider et al., 2005). Millipore's CXCR1 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 CXCR1 interaction with IL-8. The membrane preparations exhibit a Kd of 0.2 nM for [125I]-IL-8. With 5 μg/well CXCR1 Membrane Prep and 0.1 nM [125I]-IL-8, a greater than 5-fold signal-to-background ratio is obtained.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Applications
      ApplicationHuman CXCR1 / IL-8a GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay, and GTPgammaS binding.
      Biological Information
      Concentration1 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a member of the G-protein-coupled receptor family. This protein is a receptor for interleukin 8 (IL8). It binds to IL8 with high affinity, and transduces the signal through a G-protein activated second messenger system. Knockout studies in mice suggested that this protein inhibits embryonic oligodendrocyte precursor migration in developing spinal cord. This gene, IL8RB, a gene encoding another high affinity IL8 receptor, as well as IL8RBP, a pseudogene of IL8RB, form a gene cluster in a region mapped to chromosome 2q33-q36.
      Gene Symbol
      • IL8RA
      • CMKAR1
      • CKR-1
      • C-C
      • C-C-CKR-1
      • CXCR-1
      • IL8RBA
      • IL8R1
      • CDw128a
      • CD128
      • CXCR1
      • CD181
      GPCR ClassA
      Host CellsChem-1
      Protein TargetCXCR1 / IL-8a
      Target Sub-FamilyChemokine
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P25024 # Receptor to interleukin-8, which is a powerful neutrophils chemotactic factor. Binding of IL-8 to the receptor causes activation of neutrophils. This response is mediated via a G-protein that activate a phosphatidylinositol-calcium second messenger system. This receptor binds to IL-8 with a high affinity and to MGSA (GRO) with a low affinity.
      SIZE: 350 amino acids; 39791 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P25024 ## 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] Interleukin-8 (Perkin Elmer #NEX277)

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

      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 IL-8 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 varying amounts of CXCR1 Receptor membrane preparation in a dose response curve with unlabeled IL-8.
      Product Usage Statements
      5 µg/well 10 µg/well
      Signal:Background 14.8 12.8
      Specific Binding (cpm) 8965 9624


      SPECIFICATIONS: 1 unit = 5 μg

      Bmax: 3.7 pmol/mg;

      Kd: 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

      Documentation

      Protocols

      Title
      Assembly of a Competition Binding Curve

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CXCR1 RECEPTOR - 2218176 2218176

      References

      Reference overviewPub Med ID
      Reversing systemic inflammatory response syndrome with chemokine receptor pepducins.
      Kaneider NC et al.
      Nat Med., 11(6):661-5 (2005) 2005

      15880119 15880119
      Noncompetitive allosteric inhibitors of the inflammatory chemokine receptors CXCR1 and CXCR2: prevention of reperfusion injury.
      Bertini R et al.
      Proc. Natl. Acad. Sci. U S A., 101(32):11791-6 (2004) 2004

      15282370 15282370
      Cutting edge: expression of chemokine receptor CXCR1 on human effector CD8+ T cells.
      Takata H et al.
      J. Immunol., 173(4):2231-5 (2004) 2004

      15294933 15294933
      Expression and function of the chemokine receptors CXCR1 and CXCR2 in sepsis.
      Cummings CJ et al.
      J. Immunol., 162(4):2341-6 (1999) 1999

      9973513 9973513

      Data Sheet

      Title
      ChemiSCREEN CXCR1 Membrane Preparation - Data Sheet

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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