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HTS155M ChemiSCREEN™ Membrane Preparation Recombinant Human MC5 Melanocortin Receptor

Human MC5 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P33032 Entrez Gene Number: NM_005913.1
HTS155M
200 units  
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1MC5
      Description
      Catalogue NumberHTS155M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human MC5 Melanocortin Receptor
      OverviewFull-length human MC5R cDNA encoding MC5
      Background InformationThe melanocortin system consists of five seven-transmembrane spanning G-protein coupled receptors (MC1-MC5). Among the five members of the melanocortin receptor family, MC2 and MC5 are expressed in peripheral tissues. The MC2 receptor (ACTH receptor) is almost exclusively expressed in the adrenal cortex whereas MC5 is expressed in several organs including the adrenal cortex. Studies have shown that targeted disruption of the MC5R gene produced mice with a severe defect in water repulsion and thermoregulation caused by decreased production of sebaceous lipids (Chen et al., 1997). Data show a requirement for the MC5 in multiple exocrine glands for the production of numerous products, indicative of a coordinated system for regulation of exocrine gland function by melanocortin peptides. (Entwistle et al.,1990). Millipore's MC5 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 MC5. The membrane preparations exhibit a Kd of 1.3nM for [125I]-[Nle4, D-Phe7]-α-MSH (NDP-αMSH). With 0.3 nM [125I]-NDP-αMSH, 10µg/well MC5 Membrane Prep typically yields greater than 50-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, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80°C
      Quality LevelMQ100
      Applications
      ApplicationHuman MC5 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
      • MC5R
      • MC5-R
      • MC-2
      GPCR ClassA
      Host CellsChem-1
      Protein TargetMC5
      Target Sub-FamilyMelanocortin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P33032 # Receptor for MSH (alpha, beta and gamma) and ACTH. The activity of this receptor is mediated by G proteins which activate adenylate cyclase. This receptor is a possible mediator of the immunomodulation properties of melanocortins.
      SIZE: 325 amino acids; 36601 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in the brain but not in the melanoma cells.
      SIMILARITY: SwissProt: P33032 ## Belongs to the G-protein coupled receptor 1 family.
      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 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]-NDP-αMSH (Perkin Elmer#: NEX-352)
      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 50-fold signal:background with 125I labeled NDP-αMSH at 0.3 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 MC5 Receptor membrane prep.
      Product Usage Statements
      10 µg/well
      Signal:Background 147.3
      Specific Binding (cpm) 15208.7


      SPECIFICATIONS: 1 unit = 10 µg
      Bmax for [125I]-NDP-αMSH binding: 4.3 pmol/mg protein
      Kd for [125I]-NDP-αMSH binding: ~1.3 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
      HTS155M 04053252368240

      Documentation

      ChemiSCREEN™ Membrane Preparation Recombinant Human MC5 Melanocortin Receptor Analysenzertifikate

      TitelChargennummer
      CHEMISCREEN™ MEMBRANE PREPARATION RECOMBINANT HUMAN MC5 MELANOCORTIN RECEPTOR - 2349732 2349732

      Literatur

      ÜbersichtPub Med ID
      Exocrine gland dysfunction in MC5-R-deficient mice: evidence for coordinated regulation of exocrine gland function by melanocortin peptides
      Chen, W, et al
      Cell, 91:789-98 (1997)  1997

      		9413988 9413988
      Characterization of functional melanotropin receptors in lacrimal glands of the rat
      Entwistle, M L, et al
      Peptides, 11:477-83 ()  1990

      		2166277 2166277

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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