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HTS180M ChemiSCREEN™ Membrane Preparation Recombinant Human NPBW1 Neuropeptide B/W Receptor

Human NPBW1 / GPR7 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P48145 Entrez Gene Number: NM_005285.3
HTS180M
200 units  
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      Aperçu

      Tableau de caractéristiques principal

      SpeciesHost CellsProtein Target
      Human Chem-1NPBW1 / GPR7
      Description
      Catalogue NumberHTS180M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human NPBW1 Neuropeptide B/W Receptor
      OverviewFull length human GPR7 cDNA encoding NPBW1
      Background InformationNeuropeptide B (NPB) and neuropeptide W (NPW) are members of a recently identified neuropeptide family that are ligands for two highly similar receptors NPBW1 and NPBW2, both of which are coupled to Gi/o protein to inhibit intracellular cAMP production. Highest expression of NPBW1 mRNA and protein was identified in the amygdala and hypothalamic nuclei. Physiological studies demonstrate that intracerebroventricular infusion of NPBW1 ligands modulates feeding behavior, regulates the release of corticosterone, prolactin and growth hormone while also modulating pain pathway (Singh and Davenport, 2006). NPBW1 knock out male mice have shown mild adult-onset obesity and decreased locomotor activity. They become progressively hyperglycaemic and hyperinsulinaemic (Ishii et al., 2003). Millipore's NPBW1 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 NPBW1 receptor interactions with its ligand. The membrane preparations exhibit a Kd of 0.53nM for [125I]-Neuropeptide W23. With 5 ug/well NPBW1 Membrane Prep and 0.35nM [125I]-Neuropeptide W23, 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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
      Quality LevelMQ100
      Applications
      ApplicationHuman NPBW1 / GPR7 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
      • NPBWR1
      • GPR7
      • MGC129755
      GPCR ClassA
      Host CellsChem-1
      Protein TargetNPBW1 / GPR7
      Target Sub-FamilyNeuropeptide B/W
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P48145 # 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. Has a higher affinity for neuropeptide B.
      SIZE: 328 amino acids; 36117 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Found in cerebellum and frontal cortex. Detected at high levels in hipppocampus, amygdala and trachea; at moderate levels in fetal brain, pituitary gland and prostate. Not in caudate, accumbens, kidney or liver. Also detected at high levels in lung carcinoma.
      SIMILARITY: SwissProt: P48145 ## 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.
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with 5 µg/well of NPBW1 Receptor membrane prep.
      Product Usage Statements
      5µg/well
      Signal:Background 31.8
      Specific Binding (cpm) 21932


      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax: 8.74pmol/mg
      Kd: 0.53 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
      Global Trade Item Number
      Référence GTIN
      HTS180M 04053252326301