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HTS004M ChemiSCREEN™ CXCR4 Membrane Preparation

Human CXCR4 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiSCREEN™ CXCR4 Membrane Preparation: Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.
Host Cell: Chem-1 Uni Prot Number: P61073 Entrez Gene Number: NM_001008540.1, NM_003467.2
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HTS004M
200 units  
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      Descripción

      Replacement Information

      Tabla espec. clave

      SpeciesHost CellsProtein Target
      Human Chem-1CXCR4
      Description
      Catalogue NumberHTS004M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ CXCR4 Membrane Preparation
      OverviewFull-length human CXCR4 cDNA
      Background InformationThe chemokine SDF-1a and its GPCR receptor CXCR4 have a one-to-one specificity that is unique among chemokines and their receptors. SDF-1a binds to CXCR4 expressed on hematopoietic and lymphopoietic cells, and directs their trafficking to and retention in hemato-and lymphatopoietic organs and sites of inflammation (Kucia et al., 2004). CXCR4 is expressed on several tumor cell lineages, and might be responsible for metastasis to sites of SDF-1a expression, such as bone and lymph nodes (Muller et al., 2001). In addition, CXCR4 is a coreceptor for the HIV envelope glycoprotein gp120 (Feng et al., 1996). Small molecule antagonists of CXCR4 have been developed and shown to inhibit infectivity of T-tropic HIV strains and to impair growth of brain tumors (Arakaki et al., 1999; Rubin et al., 2003). Chemicon's CXCR4 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 tools for screening for antagonists of interactions between CXCR4 and SDF-1a. Radiolabeled SDF-1a binds to the CXCR4 membranes with a Kd of 0.9 nM. With 0.04-0.13 nM 125I-labeled SDF-1a, an approximately 3-fold window of specific binding is obtained.
      References
      Product Information
      FormatMembranes
      PresentationPackaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA

      Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman CXCR4 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
      Concentration2 mL, 0.5 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a CXC chemokine receptor specific for stromal cell-derived factor-1. The protein has 7 transmembrane regions and is located on the cell surface. It acts with the CD4 protein to support HIV entry into cells and is also highly expressed in breast cancer cells. Mutations in this gene have been associated with WHIM (warts, hypogammaglobulinemia, infections, and myelokathexis) syndrome. Alternate transcriptional splice variants, encoding different isoforms, have been characterized.
      Gene Symbol
      • CXCR4
      • WHIM
      • NPY3R
      • NPYY3R
      • FB22
      • LAP3
      • Fusin
      • CXCR-4
      • LCR1
      • NPYR
      • HM89
      • fusin
      • HSY3RR
      • CXC-R4
      • NPYY3
      • NPYRL
      • D2S201E
      • CD184
      • LESTR
      GPCR ClassA
      Host CellsChem-1
      Protein TargetCXCR4
      Target Sub-FamilyChemokine
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P61073 # Receptor for the C-X-C chemokine CXCL12/SDF-1. Transduces a signal by increasing the intracellular calcium ions level. Involved in haematopoiesis and in cardiac ventricular septum formation. Plays also an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. Could be involved in cerebellar development. In the CNS, could mediate hippocampal-neuron survival. Acts as a coreceptor (CD4 being the primary receptor) for HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates. Promotes Env-mediated fusion of the virus.
      SIZE: 352 amino acids; 39746 Da
      SUBUNIT: Interacts with HIV-1 surface protein gp120 and Tat.
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in numerous tissues, such as peripheral blood leukocytes, spleen, thymus, spinal cord, heart, placenta, lung, liver, skeletal muscle, kidney, pancreas, cerebellum, cerebral cortex and medulla (in microglia as well as in astrocytes), brain microvascular, coronary artery and umbilical cord endothelial cells. Isoform 1 is predominant in all tissues tested.
      DOMAIN: SwissProt: P61073 The amino-terminus is critical for ligand binding. Residues in all four extracellular regions contribute to HIV-1 coreceptor activity.
      PTM: Sulfated.
      DISEASE: SwissProt: P61073 # Defects in CXCR4 are a cause of WHIM syndrome [MIM:193670]; also called warts, hypogammaglobulinemia, infections, and myelokathexis. WHIM syndrome is an immunodeficiency disease characterized by neutropenia, hypogammaglobulinemia and extensive human papillomavirus (HPV) infection. Despite the peripheral neutropenia, bone marrow aspirates from affected individuals contain abundant mature myeloid cells, a condition termed myelokathexis.
      SIMILARITY: 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 50 mM 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] SDF-1alpha (PerkinElmer#: NEX346)

      Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM 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 4-fold signal:background with 125I-labeled SDF-1alpha at 0.13 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 CXCR4 membrane prep, 0.13 nM 125I-SDF-1α and a dilution series of unlabeled SDF-1α.
      Product Usage Statements
      5 µg/well
      Signal:Background 4.9
      Specific Binding (cpm) 9004


      Specifications:
      1 unit = 5 μg
      Bmax [125I]-SDF-1alpha binding: 1.6 pmol/mg protein
      Kd for [125I] SDF-1alpha binding: ~ 0.9 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
      Número de referencia GTIN
      HTS004M 04053252285721

      Documentation

      Protocols

      Title
      Assembly of a Competition Binding Curve

      ChemiSCREEN™ CXCR4 Membrane Preparation Ficha datos de seguridad (MSDS)

      Título

      Ficha técnica de seguridad del material (MSDS) 

      ChemiSCREEN™ CXCR4 Membrane Preparation Certificados de análisis

      CargoNúmero de lote
      ChemiscreenTM MEMBRANE PREPARATION HUMAN RECOMBINANT CXCR4 CHEMOKINE RECEPTOR - 2299342 2299342

      Referencias bibliográficas

      Visión general referenciasPub Med ID
      HIV-1 entry cofactor: functional cDNA cloning of a seven-transmembrane, G protein-coupled receptor.
      Feng, Y, et al.
      Science, 272: 872-7 (1996)  1996

      Mostrar resumen       		8629022 8629022

      Folleto

      Cargo
      Product Selection Guide - Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.

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      Categorías

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