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HTS106M ChemiSCREEN™ Membrane Preparation Recombinant Human MT2 Melatonin Receptor

Human MT2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P49286 Entrez Gene Number: NM_005959.3
HTS106M
200 units  
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1MT2
      Description
      Catalogue NumberHTS106M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human MT2 Melatonin Receptor
      OverviewFull-length human MTNR1B cDNA encoding MT2
      Background InformationMelatonin (5-methoxy-acetyltryptamine) is a hormone synthesized and secreted in a circadian manner with high levels occurring in all species at night. Melatonin has been implicated in the regulation of various neural and endocrine processes that are cued by the daily change in photoperiod (Masana and Dubocovich, 2001) Two mammalian melatonin receptor subtypes, MT1 and MT2 have been cloned. Both receptors are members of the superfamily of putative seven transmembrane G-protein coupled receptors, which are coupled to the inhibition of cyclic AMP synthesis upon agonist binding. Pharmacological studies suggest that the phase shifting effect of melatonin on circadian rhythms may be mediated by the MT2 receptor subtype (Dubocovich et al. 1998). Melatonin inhibition of dopamine release from the retina is also mediated by the MT2 receptor subtype (Dubocovich et al. 1997). Chemicon's MT2 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 at MT2. The membrane preparations exhibit a Kd of 0.1 nM for [125I]-Iodomelatonin. With 0.2 nM [125I]-Iodomelatonin, 5 µg/well MT2 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 1 mg/mL in packaging buffer, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman MT2 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
      Entrez Gene SummaryThis gene encodes one of two high affinity forms of a receptor for melatonin, the primary hormone secreted by the pineal gland. This gene product is an integral membrane protein that is a G-protein coupled, 7-transmembrane receptor. It is found primarily in the retina and brain although this detection requires RT-PCR. It is thought to participate in light-dependent functions in the retina and may be involved in the neurobiological effects of melatonin.
      Gene Symbol
      • MTNR1B
      • Mel-1B-R
      • MT2
      • MEL-1B-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetMT2
      Target Sub-FamilyMelatonin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P49286 # High affinity receptor for melatonin. Likely to mediates the reproductive and circadian actions of melatonin. The activity of this receptor is mediated by pertussis toxin sensitive G proteins that inhibit adenylate cyclase activity.
      SIZE: 362 amino acids; 40188 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in retina and less in brain and hippocampus.
      SIMILARITY: SwissProt: P49286 ## Belongs to the G-protein coupled receptor 1 family.
      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 50mM HEPES, pH 7.4, 500mM NaCl. 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, filtered and stored at 4°C
      Radioligand: [125I] Iodomelatonin (Perkin Elmer#: NEX-236)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 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 6-fold signal:background with [125I]-labeled Iodomelatonin at 0.2 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 MT2 membrane prep and unlabeled melatonin.
      Product Usage Statements
      5 µg/well
      Signal:Background 9.96
      Specific Binding (cpm) 1424.1

      SPECIFICATIONS: 1 unit = 5 µg
      Bmax for [125I]-Iodomelatonin binding: 0.04 pmol/mg protein
      Kd for [125I]-Iodomelatonin binding: ~0.1 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
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Bestellnummer GTIN
      HTS106M 04053252735462

      Documentation

      Literatur

      ÜbersichtPub Med ID
      Melatonin receptor signaling: finding the path through the dark
      Masana, M I and Dubocovich, M L
      Sci STKE, 2001:PE39 (2001)  2001

      		11698691 11698691
      Selective MT2 melatonin receptor antagonists block melatonin-mediated phase advances of circadian rhythms
      Dubocovich, M L, et al
      FASEB J, 12:1211-20 (1998)  1998

      		9737724 9737724
      Melatonin receptor antagonists that differentiate between the human Mel1a and Mel1b recombinant subtypes are used to assess the pharmacological profile of the rabbit retina ML1 presynaptic heteroreceptor
      Dubocovich, M L, et al
      Naunyn Schmiedebergs Arch Pharmacol, 355:365-75 (1997)  1997

      		9089668 9089668

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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