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HTS158M ChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor

Human alpha1B 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: P35368 Entrez Gene Number: NM_000679.3
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HTS158M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1alpha1B
      Description
      Catalogue NumberHTS158M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor
      OverviewFull-length human ADRA1B cDNA encoding α1B adrenergic
      Background InformationThe endogenous catecholamines epinephrine and norepinephrine have profound effects on smooth muscle activity, cardiac function, carbohydrate and fat metabolism, hormone secretion, neurotransmitter release, and central nervous system actions. These activities are mediated by GPCRs belonging to two subfamilies, the α- and β-adrenoceptors (Bylund et al., 1994). The three members of the α1 subclass of adrenoceptors, α1A, α1B and α1D, couple to Gq, and promote contraction of vascular and urinary tract smooth muscle, relaxation of intestinal smooth muscle, increased contractile force in the heart, and glycogenolysis and gluconeogenesis in the liver. The different subtypes have overlapping distributions and variably contribute to these effects depending on species and tissue. Overexpression of a constitutively active α1B mutant in the heart of transgenic mice resulted in cardiac hypertrophy with increased heart weight/body weight ratios. Analysis of α1B knock out mice has provided evidence that α1B is a mediator of blood pressure and aortic contractile responses induced by α1 agonists (Milano et al., 1994). The locomotor and rewarding effects of pysochostimulants and opiates were suppressed in mice lacking α1B-adrenergic receptors (Drouin et al. 2002). Millipore's α1B 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 α1B. The membrane preparations exhibit a Kd of 0.8 nM for [3H]-Prazosin. With 1 nM [3H]-Prazosin, 5μg/well α1B Membrane Prep typically yields greater than 5-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 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C
      Quality LevelMQ100
      Applications
      ApplicationHuman alpha1B 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 SummaryAlpha-1-adrenergic receptors (alpha-1-ARs) are members of the G protein-coupled receptor superfamily. They activate mitogenic responses and regulate growth and proliferation of many cells. There are 3 alpha-1-AR subtypes: alpha-1A, -1B and -1D, all of which signal through the Gq/11 family of G-proteins and different subtypes show different patterns of activation. This gene encodes alpha-1B-adrenergic receptor, which induces neoplastic transformation when transfected into NIH 3T3 fibroblasts and other cell lines. Thus, this normal cellular gene is identified as a protooncogene. This gene comprises 2 exons and a single large intron of at least 20 kb that interrupts the coding region.
      Gene Symbol
      • ADRA1B
      • ALPHA1BAR
      • ADRA1
      GPCR ClassA
      Host CellsChem-1
      Protein Targetalpha1B
      Target Sub-FamilyAdrenergic
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P35368 # This alpha-adrenergic receptor mediates its action by association with G proteins that activate a phosphatidylinositol- calcium second messenger system.
      SIZE: 520 amino acids; 56836 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P35368 ## 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 Tris, pH 7.4. 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: [3H]-Prazosin. (Perkin Elmer # NET823)

      Wash Buffer: 50 mM Tris, pH 7.4, 500mM NaCl . 0.1% BSA 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 5-fold signal:background with 3H labeled Prazosin at 1nM
      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 varying amounts of α1B Receptor membrane prep.
      Product Usage Statements
      5 µg/well
      Signal:Background 11.8
      Specific Binding (cpm) 1171

      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax for [3H]-Prazosin binding: 12.9 pmol/mg protein
      Kd ffor [3H]-Prazosin binding: ~0.8 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
      Catalogue Number GTIN
      HTS158M 04053252328619

      Documentation

      ChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN a1B ADRENERGIC RECEPTOR - 2186725 2186725

      References

      Reference overviewPub Med ID
      Decreased blood pressure response in mice deficient of the alpha1b-adrenergic receptor.
      Cavalli, A, et al.
      Proc. Natl. Acad. Sci. U.S.A., 94: 11589-94 (1997)  1997

      		9326654 9326654
      Myocardial expression of a constitutively active alpha 1B-adrenergic receptor in transgenic mice induces cardiac hypertrophy.
      Milano, C A, et al.
      Proc. Natl. Acad. Sci. U.S.A., 91: 10109-13 (1994)  1994

      		7937846 7937846

      Data Sheet

      Title
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN á1B ADRENERGIC RECEPTOR

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