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HTS110M ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT4B Serotonin Receptor

Human 5-HT4 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: Q8IXH9 Entrez Gene Number: NM_000870
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HTS110M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-15-HT4
      Description
      Catalogue NumberHTS110M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT4B Serotonin Receptor
      OverviewHuman HTR4, transcript b, encoding 5-HT4B
      Background Information5-Hydroxytryptamine (5-HT, also commonly known as serotonin) is synthesized in enterochromaffin cells in the intestine and in serotonergic nerve terminals. In the periphery, 5-HT mediates gastrointestinal motility, platelet aggregation, and contraction of blood vessels. Many functions of the central nervous system are influenced by 5-HT, including sleep, motor activity, sensory perception, arousal and appetite. A family of 12 GPCRs and one ion channel mediate the biological effects of 5-HT (Hoyer et al., 1994). 5-HT4 comprises at least 8 isoforms varying at the C-terminus, which are generated by alternative splicing. The expression and distribution of these splice variants differs among organs and tissues with many of them present in several tissues such as atrium, brain, and GI tract (Bockaert et al. 2004). To date, all isoforms have been shown to activate adenylyl cyclase in vitro, and no difference in signal transduction between C-terminal 5-HT4 receptor variants has been demonstrated. 5-HT4 receptors are of potential interest for the treatment of patients with GI motility disorders and Alzheimer's disease (De Maeyer et al. 2008; Lezoualc’h 2007). Millipore's 5-HT4B 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 5-HT4B. The membrane preparations exhibit a Kd of 0.46 nM for [3H]-GR113808. With 1 nM [3H]-GR113808, 5 μg/well 5-HT4B Membrane Prep yields greater than 10 fold signal-to-background ratio.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
      Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman 5-HT4 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
      • HTR4B
      GPCR ClassA
      Host CellsChem-1
      Protein Target5-HT4
      Target Sub-FamilySerotonin
      UniProt Number
      Incubation ConditionsRECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in assay 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 assay buffer. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with assay buffer. The plate is dried and counted.

      Assay buffer: 25 mM Tris, pH 7.4, filtered and stored at 4°C

      Radioligand: [3H]-GR113808 (Amersham TRK944)

      One package contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 10-fold signal:background with 3H-labeled GR113808 at 1 nM.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT4B membrane prep.
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 26.4 21.7
      Specific Binding (cpm) 5286 4879

      SPECIFICATIONS: 1 unit = 5 μg membrane preparation
      Bmax: 12.5 pmol/mg
      Kd: 2.7 nM for [3H]-GR113808
      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
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      HTS110M 04053252287169

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      Categories

      Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations