HTS030M | ChemiSCREEN™ PTH Membrane Preparation

HTS030M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      HumanChem-1PTH1
      Description
      Catalogue NumberHTS030M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ PTH Membrane Preparation
      OverviewFull-length human PTH1R cDNA
      Background InformationParathyroid hormone (PTH) plays a critical role in mineral ion homeostasis, particularly in bone and kidney (Mannstadt et al., 1999; Gensure et al., 2005). A related peptide, parathyroid hormone-related peptide (PTHrP), plays an important role in skeletal development. Both peptides exert their biological actions by binding to a class B GPCR, PTH1R, that signals primarily by activation of adenylate cyclase (Schipani et al., 1993). Mutations in PTH1R have been identified as the cause of Jansen's metaphyseal chondrodysplasia, Blomstrand's chondrodysplasia, and enchondromatosis (Schipani and Provot, 2003). Intermittant treatment with PTH has important clinical utility in building bone mass in patients with osteoporosis (Rosen, 2003). Chemicon's cloned human PTH1R-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant PTH1R expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between PTH1R and its ligands.
      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.
      Applications
      ApplicationHuman PTH1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay, and GTPgammaS binding.
      Biological Information
      Concentration0.5 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a member of the G-protein coupled receptor family 2. This protein is a receptor for parathyroid hormone (PTH) and for parathyroid hormone-like hormone (PTHLH). The activity of this receptor is mediated by G proteins which activate adenylyl cyclase and also a phosphatidylinositol-calcium second messenger system. Defects in this receptor are known to be the cause of Jansen's metaphyseal chondrodysplasia (JMC), chondrodysplasia Blomstrand type (BOCD), as well as enchodromatosis.
      Gene Symbol
      • PTHR1
      • PTHR
      • MGC138426
      • MGC138452
      GPCR ClassB
      Host CellsChem-1
      Protein TargetPTH1
      Target Sub-FamilyPTH receptor
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q03431 # This is a receptor for parathyroid hormone and for parathyroid hormone-related peptide. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase and also a phosphatidylinositol-calcium second messenger system.
      SIZE: 593 amino acids; 66361 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in most tissues. Most abundant in kidney, bone and liver.
      DISEASE: SwissProt: Q03431 # Defects in PTHR1 are the cause of Jansen metaphyseal chondrodysplasia (JMC) [MIM:156400]. JMC is a rare autosomal dominant disorder characterized by a short-limbed dwarfism associated with hypercalcemia and normal or low serum concentrations of the two parathyroid hormones. & Defects in PTHR1 are the cause of chondrodysplasia Blomstrand type (BOCD) [MIM:215045]. BOCD is a severe skeletal dysplasia. & Defects in PTHR1 may be a cause of enchondromatosis [MIM:166000]. Enchondromas are common benign cartilage tumors of bone. They can occur as solitary lesions or as multiple lesions in enchondromatosis (Ollier and Maffucci diseases). Clinical problems caused by enchondromas include skeletal deformity and the potential for malignant change to osteosarcoma.
      SIMILARITY: SwissProt: Q03431 ## Belongs to the G-protein coupled receptor 2 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, 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] PTH (Perkin Elmer#: NEX397)

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

      One vial 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 125I-labeled PTH at 0.75 nM
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceSPECIFICATIONS: Bmax: 2.66 pmol/mg
      Kd: 0.75 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

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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