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HTS028M ChemiSCREEN™ Membrane Preparation Recombinant Human sst2 Somatostatin Receptor

Human sst2 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: P30875 Entrez Gene Number: NM_001050.2
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HTS028M
200 units  
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      Overview

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1sst2
      Description
      Catalogue NumberHTS028M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human sst2 Somatostatin Receptor
      OverviewFull-length human SSTR2 cDNA encoding sst2
      Background InformationSomatostatin (sst) is a multifunctional peptide with two biologically active forms, sst-14 and sst-28, which are synthesized in neurons throughout the brain as well as in peripheral tissues such as the pancreas and the gut (Gillies, 1997). SST exerts a diverse array of effects that include inhibition of endocrine secretion, modulation of neurotransmission, and regulation of cell proliferation by stimulating a family of five G-protein-coupled receptors. Somatostatin receptor sst2 mRNA is predominantly expressed in central nervous system. Study using sst2 knock-out mice has found the increased anxiety-related behaviour while locomotor and exploratory activity was decreased in stress-inducing situations (coupled with an increase in pituitary ACTH release, a regulator of the stress response) (Viollet et al., 2000) . In the periphery, inhibition of glucagon release by sst in mouse islets is primarily mediated via sst2 (Strowski et al., 2000). In addition, endogenous sst functions through sst2 to suppress gastric acid secretion through inhibition of gastrin activity (Martinez et al., 1998). Millipore's sst2 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 agonists and antagonists of sst2. The membrane preparations exhibit a Kd of 0.95 nM for [125I]-Somatostatin 14. With 0.75 nM [125I]-Somatostatin 14, 5 μg/well sst2 Membrane Prep typically yields greater than 20-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 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C
      Quality LevelMQ100
      Applications
      ApplicationHuman sst2 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
      • SST2
      GPCR ClassA
      Host CellsChem-1
      Protein Targetsst2
      Target Sub-FamilySomatostatin
      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]-Somatostatin 14. (Perkin Elmer NEX-389 )

      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 20-fold signal:background with 125I labeled somatostatin 14 at 0.75 nM
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of sst2 Receptor membrane prep.
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 16.2 36.2
      Specific Binding (cpm) 58438 54632


      SPECIFICATIONS: 1 unit = 5 μg
      Bmax for [125I]-Somatostatin binding: 18.9 pmol/mg protein
      Kd for [125I]-Somatostatin binding: ~0.95 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
      Catalogue Number GTIN
      HTS028M 04053252582769