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HTS059M ChemiScreen™ V1A Vasopressin Membrane Preparation

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HTS059M
200 units  1 mL, 2 mg/mL
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1V1A
      Description
      Catalogue NumberHTS059M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ V1A Vasopressin Membrane Preparation
      OverviewHuman AVPR1 cDNA encoding V1A
      Materials Required but Not DeliveredMembranes 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] vasopressin V1A antagonist (Perkin Elmer catalog # NEX310)
      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 4-fold signal:background with 125I-labeled vasopressin V1A antagonist at 0.1 nM
      Background InformationArginine vasopressin (AVP) is a 9 amino acid peptide that functions as an antidiuretic, vasoconstrictor and neurotransmitter. The three vasopressin receptors, V1A, V1B and V2, are GPCRs; V1A and V1B couple to Gq and calcium release, whereas V2 couples to Gs. V1 receptors regulate fluid homeostasis by stimulating contraction of glomerular mesangial cells and renal vascular smooth muscle cells (Birnbaumer, 2000). Chemicon's V1A 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 V1A interactions with its ligands. The membrane preparations exhibit a Kd of 55 pM for [125I]-vasopressin antagonist ([125I]-phenylacetyl-D-Tyr(Me)-Phe-Gln-Asn-Arg -Pro-Arg-Tyr-NH2). With 10 µg/well V1A Membrane Prep and 0.1 nM [125]-vasopressin V1A antagonist, greater than 12-fold signal-to-background ratio were obtained.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives. Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman V1A 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
      Concentration0.5 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene acts as receptor for arginine vasopressin. This receptor belongs to the subfamily of G-protein coupled receptors which includes AVPR1B, V2R and OXT receptors. Its activity is mediated by G proteins which stimulate a phosphatidylinositol-calcium second messenger system. The receptor mediates cell contraction and proliferation, platelet aggregation, release of coagulation factor and glycogenolysis.
      Gene Symbol
      • AVPR1A
      • AVPR1
      • V1aR
      GPCR ClassA
      Host CellsChem-1
      Protein TargetV1A
      Target Sub-FamilyVasopressin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P37288 # Receptor for arginine vasopressin. The activity of this receptor is mediated by G proteins which activate a phosphatidyl- inositol-calcium second messenger system. Has been involved in social behaviors, including affiliation and attachment.
      SIZE: 418 amino acids; 46800 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P37288 ## Belongs to the G-protein coupled receptor 1 family.
      MISCELLANEOUS: Differences in regional receptor expression in the brain as well as differences in social behavior may result from a highly variable repetitive sequence in the 5' flanking region of AVPR1A. One such allelic variant has been linked to autism.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of V1A membrane prep:
      Product Usage Statements
      10 µg/well
      Signal:Background 24.4
      Specific Binding (cpm) 11827


      SPECIFICATIONS:
      1 unit = 10 µg
      Bmax: 1.76 pmol/mg
      Kd: 55 pM
      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
      Material Package1 mL, 2 mg/mL
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Bestellnummer GTIN
      HTS059M 04053252506598

      Documentation

      ChemiScreen™ V1A Vasopressin Membrane Preparation SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

      Literatur

      ÜbersichtPub Med ID
      Vasopressin receptors.
      Birnbaumer, M
      Trends Endocrinol. Metab., 11: 406-10 (2000)  1999

      Abstract anzeigen
      11091117 11091117

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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