HTS111M ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT6 Serotonin Receptor

HTS111M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1 5-HT6
      Description
      Catalogue NumberHTS111M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT6 Serotonin Receptor
      OverviewFull-length human HTR6 cDNA encoding the 5-HT6 Serotonin Receptor
      Background InformationThe neurotransmitter serotonin/5-hydroxytryptamine (5-HT) regulates a wide variety of neurological functions. A family of 13 receptors (12 GPCRs and one ion channel) mediate the effects of serotonin. The serotonin receptor 5-HT6 is a Gs coupled receptor expressed solely in the CNS, primarily in the limbic and cortical regions. 5-HT6 appears to play a role in memory and learning, obesity, psychosis, anxiety and epilepsy (Woolley et al., 2004; Fisas et al., 2006). In particular, a 5-HT6-selective agonist caused significant weight loss in a rat model of diet-induced obesity. Millipore's 5-HT6 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-HT6. The membrane preparations exhibit a Kd of 0.44 nM for [125I]-SB258585. With 0.25nM [125I]-SB258585, 5 μg/well 5-HT6 Membrane Prep typically yields greater than 6-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: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Applications
      ApplicationHuman 5-HT6 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
      • HTR6
      GPCR ClassA
      Host CellsChem-1
      Protein Target5-HT6
      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 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 the binding buffer. 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: 20 mM HEPES, 3 mM MgCl2, 2 mM Ascorbic acid, pH 7.4, filtered and stored at 4°C

      Radioligand: [125I]-SB258585 (Perkin Elmer #:NEX424)

      Wash Buffer: same as the binding buffer.

      One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 6-fold signal:background with 125I labeled SB258585 at 0.25 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-HT6 Receptor membrane prep.
      Product Usage Statements
      5 µg/well 10 µg/well
      Signal:Background 12.8 12.3
      Specific Binding (cpm) 10192 12675


      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax for [125I]-SB258585 binding: 6.1 pmol/mg protein
      Kd for [125I]-SB258585 binding: ~0.44 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 up to expiration date indicated on the label. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT6 Serotonin Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT6 Serotonin Receptor Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 5-HT6 SERATONIN RECEPTOR - 23244772324477

      References

      Reference overviewPub Med ID
      Chronic 5-HT6 receptor modulation by E-6837 induces hypophagia and sustained weight loss in diet-induced obese rats.
      Fisas, Angels, et al.
      Br. J. Pharmacol., 148: 973-83 (2006)  2006

      16783408 16783408
      5-ht6 receptors.
      Woolley, Marie L, et al.
      Current drug targets. CNS and neurological disorders, 3: 59-79 (2004)  2004

      Show Abstract
      14965245 14965245

      Data Sheet

      Title
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 5-HT6 SERATONIN RECEPTOR
      CHEMISCREEN™ MEMBRANE PREPARATION HUMAN RECOMBINANT 5-HT6 SEROTONIN RECEPTOR

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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