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HTS181M ChemiSCREEN™ Membrane Preparation Recombinant Human NPBW2 Neuropeptide B/W Receptor

Human NPBW2 / GPR8 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P48146 Entrez Gene Number: NM_005286.2
HTS181M
200 units  
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1NPBW2 / GPR8
      Description
      Catalogue NumberHTS181M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human NPBW2 Neuropeptide B/W Receptor
      OverviewFull length human GPR8 cDNA encoding NPBW2
      Background InformationNeuropeptide B (NPB) and neuropeptide W (NPW) are members of a recently identified neuropeptide family that are ligands for NPBW1 and NPBW2 receptors, both of which are coupled to Gi/o protein to inhibit intracellular cAMP production and share 64% sequence homology (Singh and Davenport, 2006). NPBW1 recognizes both NPB and NPW with similar nanomolar affinities (with a slight preference for NPB), whereas NPBW2 is moderately selective for NPW (Tanaka et al., 2003). NPBW2 is one of a few GPCRs that have no rat or mouse orthologue, although gene encoding NPBW2 has been discovered in other mammalian species such as rabbit (Lee et al., 1999). NPBW2 mRNA is known to be expressed in the frontal cortex, parietal cortex hippocampus, caudate nucleus, thalamus, pituitary, adrenal gland, and lymph node (Brezillon et al., 2003). Functions of NPBW2 may be involved in feeding, weight regulation, and pain response through direct or indirect actions in the central nervous system. Millipore's NPBW2 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 NPBW2 receptor interactions with its ligand. The membrane preparations exhibit a Kd of 2nM for [125I]-Neuropeptide W23. With 10 ug/well NPBW2 Membrane Prep and 0.35nM [125I]-Neuropeptide W23, a greater than 10-fold signal-to-background ratio was obtained.
      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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
      Quality LevelMQ100
      Applications
      ApplicationHuman NPBW2 / GPR8 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
      Entrez Gene SummaryThe protein encoded by this gene is an integral membrane protein and G protein-coupled receptor. The encoded protein is similar in sequence to another G protein-coupled receptor (GPR7), and it is structurally similar to opioid and somatostatin receptors. This protein binds neuropeptides B and W. This gene is intronless and is expressed primarily in the frontal cortex of the brain.
      Gene Symbol
      • NPBWR2
      • GPR8
      GPCR ClassA
      Host CellsChem-1
      Protein TargetNPBW2 / GPR8
      Target Sub-FamilyNeuropeptide B/W
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P48146 # Interacts specifically with a number of opioid ligands. Receptor for neuropeptides B and W, which may be involved in neuroendocrine system regulation, food intake and the organization of other signals.
      SIZE: 333 amino acids; 36861 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Detected at high levels in caudate nucleus, hyppocampus and amygdala; at moderate levels in the adult brain, thalamus, parietal cortex, pituitary gland, adrenal gland and lymph nodes.
      SIMILARITY: SwissProt: P48146 ## Belongs to the G-protein coupled receptor 1 family.
      Incubation ConditionsMembranes 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 Tris, pH 7.4. 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, filtered and stored at 4°C.
      Radioligand: [125I]-neuropeptide W23 (Perkin Elmer#: NET-403)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 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 neuropeptide W23 at 0.35nM.
      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 10 or 20 µg/well of NPBW2 Receptor membrane prep.
      Product Usage Statements
      20 µg/well 10 µg/well
      Signal:Background 26.3 30.2
      Specific Binding (cpm) 20148 19282


      SPECIFICATIONS: 1 unit = 10.0 µg membrane preparation
      Bmax: 11.34 pmol/mg
      Kd: 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
      HTS181M 04053252672699

      Documentation

      Literatur

      ÜbersichtPub Med ID
      Neuropeptide B and W: neurotransmitters in an emerging G-protein-coupled receptor system.
      Singh, Gurminder and Davenport, Anthony P
      Br. J. Pharmacol., 148: 1033-41 (2006)  2005

      Abstract anzeigen       		16847439 16847439
      Identification of natural ligands for the orphan G protein-coupled receptors GPR7 and GPR8
      Brezillon, Stephane, et al
      J Biol Chem, 278:776-83 (2003)  2003

      		12401809 12401809
      Characterization of a family of endogenous neuropeptide ligands for the G protein-coupled receptors GPR7 and GPR8
      Tanaka, Hirokazu, et al
      Proc Natl Acad Sci USA, 100:6251-6 (2003)  2003

      		12719537 12719537

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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