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HTS066M ChemiSCREEN™ Human Y2 Neuropeptide Receptor Membrane Preparation

Human Y2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P49146 Entrez Gene Number: NM_000910.2
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HTS066M
200 units  
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      Aperçu

      Tableau de caractéristiques principal

      SpeciesHost CellsProtein Target
      Human Chem-1Y2
      Description
      Catalogue NumberHTS066M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Human Y2 Neuropeptide Receptor Membrane Preparation
      OverviewFull-length human NPY2R cDNA encoding Y2
      Background InformationThe NPY family consists of three 36-amino acid peptides, neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP), which bind to the NPY receptor family of G protein-coupled receptors. Five NPY receptors, Y1, Y2, Y4, Y5 and Y6, have been defined at the molecular level, and each signals primarily through Gi/o. Binding of NPY family peptides to NPY receptors mediates a variety of physiological effects, including promotion of food intake, decreased anxiety, inhibition of neurotransmitter and hormone release, vasoconstriction, and gut motility. Y2 is primarily expressed in the CNS, and it mediates presynaptic inhibition of neurotransmitter release (Michel et al., 1998). Chemicon's Y2 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 Y2 interactions and its ligands. The membrane preparations exhibit a Kd of 1.28 nM for [125I]-human PYY. With 0.5 nM [125I]-human PYY, 5 or 10µg/well Y2 Membrane Prep yields greater than 10-fold signal-to-background ratio.
      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 0.5 mg/ml in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman Y2 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
      • NPY2R
      • NPY2-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetY2
      Target Sub-FamilyNeuropeptide Y
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P49146 # Receptor for neuropeptide Y and peptide YY. The rank order of affinity of this receptor for pancreatic polypeptides is PYY > NPY > PYY (3-36) > NPY (2-36) > [Ile-31, Gln-34] PP > [Leu- 31, Pro-34] NPY > PP, [Pro-34] PYY and NPY free acid.
      SIZE: 381 amino acids; 42731 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: High levels in amygdala, corpus callosum, hippocampus and subthalamic nucleus. Also detectable in caudate nucleus, hypothalamus and substantia nigra.
      SIMILARITY: SwissProt: P49146 ## 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, a GF/C 96-well filter plate 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]-human PYY (Perkin Elmer#:NEX-341 )
      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 a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I labeled human PYY at 0.5 nM
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of TP membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 19.8 26.3
      Specific Binding (cpm) 27347 23335


      SPECIFICATIONS: 1 unit = 5 µg
      Bmax for [125I] human PYY binding: 17.89 pmol/mg protein;
      Kd for [125I] human PYY binding: ~ 1.28 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
      HTS066M 04053252510090