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HTS215M ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor

Human 5-HT7 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: P34969 Entrez Gene Number: NM_000872
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HTS215M
200 units  
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      Overview

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-15-HT7
      Description
      Catalogue NumberHTS215M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor
      OverviewFull-length human HTR7 cDNA encoding the 5-HT₇ 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-HT₇ is a Gs coupled receptor expressed in the brain (hypothalamus, hippocampus and frontal cortex) and in smooth muscle and GI tract. Systemic delivery of serotonin results in hypothermia, and pharmacological and genetic studies indicate that 5-HT₇ mediates this effect (Hedlund et al., 2003). In addition, 5-HT₇ appears to modulate circadian rhythms and smooth muscle relaxation, and is implicated as a target for sleep, particularly as related to depression, and migraine (Terron and Falcon-Neri, 1999; Mullins et al., 1999). Millipore's 5-HT₇ 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-HT₇. The membrane preparations exhibit a Kd of 37.7 nM for [³H]-5-Carboxamidotryptamine (5-CT). With 30 nM [³H]-5-CT, 10 μg/well 5-HT₇ Membrane Prep typically yields greater than 4-fold signal-to-background ratio.
      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.
      Quality LevelMQ100
      Applications
      ApplicationHuman 5-HT7 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
      • HTR7
      GPCR ClassA
      Host CellsChem-1
      Protein Target5-HT7
      Target Sub-FamilySerotonin
      UniProt Number
      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, 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: 50 mM Tris, pH 7.4, 5 mM MgCl₂, 4 mM CaCl₂, 0.5 mM ascorbic acid, filtered and stored at 4°C

      Radioligand: [³H]-5-Carboxamidotryptamine (Perkin Elmer NET1071)

      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 4-fold signal:background with ³H-labeled 5-CT at 30 nM
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with 10 μg of 5-HT₇ Receptor membrane prep.
      Product Usage Statements
      10 µg/well
      Signal:Background 7.5
      Specific Binding (cpm) 545

      SPECIFICATIONS: 1 unit =10 μg
      Bmax for [³H]-5-CT binding: 20.1 pmol/mg protein
      Kd for [³H]-5-CT binding: ~37.7 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
      HTS215M 04053252474309