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HTS098M ChemiSCREEN™ Membrane Preparation Recombinant Human NMU2 Neuromedin U Receptor

Human NMU2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: Q9GZQ4 Entrez Gene Number: NM_006056
HTS098M
200 units  
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      Overview

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1NMU2
      Description
      Catalogue NumberHTS098M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human NMU2 Neuromedin U Receptor
      OverviewHuman NMU2
      Background InformationNeuromedin U (NmU) is a peptide which regulates peripheral functions such as smooth muscle contraction and blood pressure, and CNS functions including nociception and feeding activity (Brighton et al., 2004a). Two GPCRs, NMU1 and NMU2, mediate the contractile effects of neuromedin U by activation of both Gq and Gi (Brighton et al., 2004b). Compared to the wide distribution of NMU1 in peripheral tissue, expression of NMU2 receptor is limited to areas of the brain, such as the paraventricular nucleus, along the wall of the third ventricle in the hypothalamus and the CA1 region of the hippocampus, and to the spinal cord (Howard et al. 2000). Recent study has shown that mice deficient in NMU2 but not NMU1 receptor had impaired nociceptive responses suggesting that the pro-nociceptive effects of NmU in mice appear to be mediated through NMU2 (Zeng et al., 2006; Torres et al. 2007). Millipore's NMU2 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 NMU1 interactions with neuromedin U25. The membrane preparations exhibit a Kd of 0.58 nM for [125I]-neuromedin U-25. With 5 μg/well NMU2 Membrane Prep and 0.25 nM [125I]-neuromedin U-25, a greater than 10-fold signal-to-background ratio was obtained.
      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 NMU2 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
      • NMUR2
      • NMU2R
      • TGR1
      GPCR ClassA
      Host CellsChem-1
      Protein TargetNMU2
      Target Sub-FamilyNeuromedin U
      UniProt Number
      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]-neuromedin U-25 (Perkin Elmer # NEX383)

      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 neuromedin U-25 at 0.25 nM.
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with NMU2 membrane prep.
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 22.4 22.5
      Specific Binding (cpm) 10666 7780

      SPECIFICATIONS: 1 unit = 5 μg membrane preparation
      Bmax: 3.92 pmol/mg
      Kd: 0.58 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
      Global Trade Item Number
      Catalogue Number GTIN
      HTS098M 04053252671425