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HTS065M ChemiSCREEN™ Membrane Preparation Recombinant Human MT1 Melatonin Receptor

Human MT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P48039 Entrez Gene Number: NM_005958.3
HTS065M
200 units  
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      Übersicht

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1MT1
      Description
      Catalogue NumberHTS065M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human MT1 Melatonin Receptor
      OverviewFull-length human MTNR1A cDNA encoding MT1
      Background InformationMelatonin (5-methoxy-acetyltryptamine) is a vertebrate hormone synthesized and secreted in a circadian manner with high levels occurring at night. Melatonin has been implicated in photoperiod regulation of various neural and endocrine processes, including seasonal reproduction, body weight, metabolism, and coat color (Masana and Dubocovich, 2001). In mammals, the melatonin rhythm is generated by an endogenous circadian clock in the suprachiasmatic nucleus (SCN) of the hypothalamus, which is entrained by the light/dark cycle to the 24h day. Melatonin may be effective in synchronizing disturbed circadian rhythms in humans, and there is considerable interest in melatonin's potential for treatment of disordered circadian rhythms in jet-lag, shift work, blindness and in delayed/advanced sleep phase syndromes (Hagan and Oakley, 1995). The two mammalian melatonin receptor subtypes, MT1 and MT2, are G-protein coupled receptors which signal through inhibition of cyclic AMP synthesis (Dubocovich et al., 2000). In addition, MT1 is involved in regulation of proliferation of human breast cancer cells (Ram et al., 2002; Collins et al., 2003). Millipore's MT1 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 MT1 interactions with its ligands. The membrane preparations exhibit a Kd of 1.7 nM for [125I]-Iodomelatonin. With 0.5 nM [125I]-Iodomelatonin, 5 µg/well MT1 Membrane Prep typically yields greater than 15-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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman MT1 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 receptor is a G-protein coupled, 7-transmembrane receptor that is responsible for melatonin effects on mammalian circadian rhythm and reproductive alterations affected by day length. The receptor is an integral membrane protein that is readily detectable and localized to two specific regions of the brain. The hypothalamic suprachiasmatic nucleus appears to be involved in circadian rhythm while the hypophysial pars tuberalis may be responsible for the reproductive effects of melatonin.
      Gene Symbol
      • MTNR1A
      • MT1
      • Mel-1A-R
      • MEL-1A-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetMT1
      Target Sub-FamilyMelatonin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P48039 # 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: 350 amino acids; 39375 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in hypophyseal pars tuberalis and hypothalamic suprachiasmatic nuclei (SCN). Hippocampus.
      SIMILARITY: SwissProt: P48039 ## 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 15-fold signal:background with 125I-labeled Iodomelatonin at 0.5 nM
      Signal:background and specific binding values obtained in a competition binding assay with MT1 membrane prep and unlabeled melatonin:
      Product Usage Statements
      10 µg/well
      Signal:Background 28.5
      Specific Binding (cpm) 104301


      SPECIFICATIONS: 1 unit = 5 µg
      Bmax for [125I]-Iodomelatonin binding: 22.8 pmol/mg protein
      Kd for [125I]-Iodomelatonin binding: ~1.7 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
      Bestellnummer GTIN
      HTS065M 04053252733970