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HTS123M ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation

Human BB1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P28336 Entrez Gene Number: NM_002511.2
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HTS123M
200 units  1 mL per vial
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1BB1
      Description
      Catalogue NumberHTS123M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation
      OverviewFull-length human NMBR cDNA encoding BB1
      Background InformationBombesin, a bioactive peptide first identified in amphibian skin, is related to two mammalian peptides, gastrin-releasing peptide (GRP) and neuromedin B (NMB). A family of 3 GPCRs, including NMB-R (BB1), GRP-R (BB2), and BRS-3 (BB3), mediate the biological effects of the peptides. The receptors differ in their affinities for the peptides; BB2 binds to GRP with 50-300-fold greater affinity than to NMB, whereas BB1 binds to NMB with 10-800-fold greater affinity than to GRP (Tokita et al., 2004). Binding of ligand to BB1 activates Gq to increase intracellular calcium concentrations. The CNS is a major site of NMB and BB1 expression, and BB1 appears to be involved in thermoregulation (Ohki-Hamazaki et al., 2005). Chemicon's BB1 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 antagonists of BB1 interactions with neuromedin B. The membrane preparations exhibit a Kd of 0.35-0.43 nM for [125I]-Bombesin. With 5 µg/well BB1 Membrane Prep and 0.3 nM [125I]-bombesin, a greater than 40-fold signal-to-background ratio was obtained.
      References
      Product Information
      FormatMembranes
      PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 40-fold signal:background with 125I-labeled bombesin at 0.3 nM.

      Liquid 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 BB1 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 SummaryNeuromedin B receptor binds neuromedin B, a potent mitogen and growth factor for normal and neoplastic lung and for gastrointestinal epithelial tissue.
      Gene Symbol
      • NMBR
      • NMB-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetBB1
      Target Sub-FamilyBombesin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P28336 # Receptor for neuromedin-B.
      SIZE: 390 amino acids; 43453 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P28336 ## 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, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. 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, 0.2% BSA, filtered and stored at 4°C
      Radioligand: [125I] bombesin (Perkin Elmer # NEX258)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of BB1 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 82.5 74.8
      Specific Binding (cpm) 27377 22443


      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax: 3.1 pmol/mg
      Kd: 0.39 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
      Material Package1 mL per vial
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Bestellnummer GTIN
      HTS123M 04053252624124

      Documentation

      ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

      ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation Analysenzertifikate

      TitelChargennummer
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN BB1 BOMBESIN/NEUROMEDIN B RECEPTOR - 2037872 2037872

      Literatur

      ÜbersichtPub Med ID
      Development and function of bombesin-like peptides and their receptors
      Ohki-Hamazaki, Hiroko, et al
      Int J Dev Biol, 49:293-300 (2005)  2004

      		15906244 15906244
      Molecular basis of the selectivity of gastrin-releasing peptide receptor for gastrin-releasing peptide
      Tokita, Kenji, et al
      Mol Pharmacol, 61:1435-43 (2002)  2002

      		12021405 12021405

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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