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HTS160M ChemiSCREEN™ Membrane Preparation Recombinant Human BB3 Bombesin Receptor

Human BB3 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: NM_001727
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HTS160M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1BB3
      Description
      Catalogue NumberHTS160M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human BB3 Bombesin Receptor
      OverviewFull length human BRS3 encoding BB3
      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 GRP-R (BB1), NMB-R (BB2) and BRS-3 (BB3), mediate the biological effects of the peptides (Ohki-Hamazaki et al., 2005). BB3 differs from the others by its low affinity for bombesin. Although an endogenous ligand for BB3 has yet to be identified, a synthetic nonselective bombesin-like peptide [H-D-Phe6, β-Ala11, Phe13, Nle14]-bombesin-(6-14)-nonapeptide amide (Bombesin (6-14) Analog) activates BB3 with high potency. BB3-null mice have an obese phenotype (Matsumoto and Iijima, 2003). Millipore's BB3 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 BB3 receptor interactions with its ligand. The membrane preparations exhibit a Kd of 0.23nM for [125I]-Bombesin (6-14) Analog. With 10 μg/well BB3 Membrane Prep and 0.3 nM [125I]-Bombesin (6-14) Analog, a greater than 3-fold signal-to-background ratio was obtained.
      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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
      Quality LevelMQ100
      Applications
      ApplicationHuman BB3 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 TargetBB3
      Target Sub-FamilyBombesin
      UniProt Number
      Incubation 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, 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, 0.2 mg/ml bacitracin, 20 μg/ml leupeptin, 20 μg/ml chymostatin, 1 Protease Inhibitor cocktail Tablets (Roche Cat. No. 11 873 580 001) for each 50 ml binding buffer.

      Radioligand: [125I]-Bombesin (6-14) Analog (Perkin Elmer NEX377)

      Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.

      One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 3-fold signal:background with 125I-labeled Bombesin (6-14) Analog 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 BB3 membrane prep.
      Product Usage Statements
      10 µg/well
      Signal:Background 5.2
      Specific Binding (cpm) 3403

      SPECIFICATIONS: 1 unit = 10 µg
      Bmax 0.95 pmol/mg protein
      Kd 0.3 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
      Catalog Number GTIN
      HTS160M 04053252619946

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