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HTS084M ChemiScreen™ BB2 Bombesin Receptor Membrane Preparation

Human BB2 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: P32247 Entrez Gene Number: M73481
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HTS084M
200 assays  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1BB2
      Description
      Catalogue NumberHTS084M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ BB2 Bombesin Receptor Membrane Preparation
      OverviewFull-length human GRPR cDNA encoding BB2
      Background InformationBombesin, a bioactive peptide first identified in amphibian skin, is related to two mammalian peptides, gastrin-releasing peptide (GRP) and neuromedin B. 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. Binding of ligand to BB2 activates Gq to increase intracellular calcium concentrations. GRP stimulates release of gastrin from endocrine cells and stimulates smooth muscle activity in the gastrointestinal tract. In addition, binding of GRP to BB2 stimulates proliferation of a variety of cell types, and has been implicated in the progression of small cell lung cancer and other malignancies. The CNS is also a major site of GRP expression, and GRP and BB2 are involved in the circadian clock, conditioned fear, and food intake (Ohki-Hamazaki et al., 2005). Chemicon's BB2 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 BB2 interactions and its ligands. The membrane preparations exhibit a Kd of 0.114 nM for [125I]-Tyr4-Bombesin. With 0.3nM [125I]-Tyr4-Bombesin, 5 µg/well or 10 µg/well BB2 Membrane Prep typically yield greater than 20-fold signal-to-background ratio.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
      Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman BB2 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
      • BRS3
      GPCR ClassA
      Host CellsChem-1
      Protein TargetBB2
      Target Sub-FamilyBombesin
      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, 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
      Radioligand: [125I]-Tyr4-Bombesin (Perkin Elmer#:NEX-258 )
      Wash Buffer: 50 mM Tris, pH 7.4 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 10-fold signal:background with 125I labeled Bombesin at 0.3 nM
      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 BB2 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 24 23
      Specific Binding (cpm) 19,260 12,610


      SPECIFICATION: Bmax: 1.28 pmol/mg; Kd: 0.114 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 assays
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      HTS084M 04053252672101

      Documentation

      ChemiScreen™ BB2 Bombesin Receptor Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub 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)  2005

      		15906244 15906244

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