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HTS081M ChemiScreen™ Membrane Preparation
Recombinant Human TBXA2R (TP) Thromboxane Receptor

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HTS081M
2 mL  200 units
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1TP / TXA2 / PGH2
      Description
      Catalogue NumberHTS081M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Membrane Preparation
      Recombinant Human TBXA2R (TP) Thromboxane Receptor
      OverviewHuman Full-length human TBXA2R cDNA encoding the TP short form (TPα)
      Background InformationTP receptors (Thromboxane A2 receptors) are widely distributed among different organ systems and have been localized on both cell membranes and intracellular structures. TP receptors belong to G protein-coupled receptor family (Hirata et al., 1991). Activation of TP receptors induces platelet aggregation, vascular and respiratory smooth muscle constriction, and enhances mitogenic responses of vascular smooth muscle cells that are stimulated by growth factors (Ali et al, 1993; Hanasaki et al., 1990). The human TP receptor is encoded by a single gene that is alternatively spliced at the carboxyl terminus, resulting in two isoforms, TPα (343 residues) and TPβ (407 residues). Both isoforms couple to Gq pathway, but couple oppositely to adenylate cyclase. The cDNA encoding human TPα has been stablely expressed in the Chem-1 host, which supports high levels of recombinant receptor expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. The membrane preparations exhibit a Kd of 9.19 nM for [3H]-SQ 29548. With 10 nM [3H]-SQ 29548, 10 ug/well TP Membrane Prep yields greater than 5-fold signal-to-background ratio.
      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 5-fold signal:background with 3H-labeled SQ 29548 at 15 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 TP / TXA2 / PGH2 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
      • TBXA2R
      • TXA2-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetTP / TXA2 / PGH2
      Target Sub-FamilyProstanoid
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P21731 # Receptor for thromboxane A2 (TXA2), a potent stimulator of platelet aggregation. The activity of this receptor is mediated by a G-protein that activate a phosphatidylinositol-calcium second messenger system. In the kidney, the binding of TXA2 to glomerular TP receptors causes intense vasoconstriction.
      SIZE: 369 amino acids; 40092 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      DISEASE:SwissProt: P21731 # Defects in TBXA2R are the cause of a dominantly inherited bleeding disorder [MIM:188070].
      SIMILARITY: SwissProt: P21731 ## 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 50 mM 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: [3H]-SQ 29548 (Perkin Elmer # NET936)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM 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 Y2 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 6.6 4.6
      Specific Binding (cpm) 2090 963


      1 unit = 10 μg membrane preparation
      Bmax: 13.9 pmol/mg
      Kd: 14.6 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 ConditionsStore at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.
      Packaging Information
      Material Size2 mL
      Material Package200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      HTS081M 04053252742927

      Documentation

      ChemiScreen™ Membrane Preparation
      Recombinant Human TBXA2R (TP) Thromboxane Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Cloning and expression of the cDNA for a human thromboxane A2 receptor gene.
      Hirata M et al.
      Nature , 349:617-620 (1991)  1991

      1825698 1825698
      Biochemical characterization and comparison of rat thromboxane A2/prostaglandin H2 receptors in platelets and cultures aortic smooth muscle cells.
      Hanasaki K., et al.
      Biochem. Pharmacol., 38:2967-2976 (1989)  1989

      2528958 2528958

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      Categories

      Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations