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HTS127M ChemiScreen™ Membrane Preparation
Recombinant Human AMY1 Calcitonin Receptor

Human AMY1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P30988 Entrez Gene Number: NM_001742.2
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HTS127M
200 units  
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1AMY1
      Description
      Catalogue NumberHTS127M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Membrane Preparation
      Recombinant Human AMY1 Calcitonin Receptor
      OverviewFull-length human CALCR (isoform 2) and RAMP1 cDNAs encoding AMY1
      Background InformationThe calcitonin family of peptides, which includes calcitonin, two calcitonin-related gene peptides (CGRPs), adrenomedullin, and amylin, share a disulfide-bonded loop, an amphipathic a-helix, and an amidated C-terminus. The peptides influence a variety of physiological effects, including vascular tone, food intake and bone metabolism. The biological effects of the peptides are mediated by two class B GPCRs, the CT and CL receptors (Poyner et al., 2002). The pharmacology of the CT and CL receptors is strongly influenced by their association with a family of three receptor activity modifying proteins (RAMPs), which are 14-17 kD single pass transmembrane proteins. When expressed with no modifying proteins, the CT receptor binds with high affinity only to calcitonin. The CT receptor in complex with RAMP proteins binds amylin with high affinity, as well as calcitonin and αCGRP; the CT receptor complexes with RAMP1, RAMP2 and RAMP3 are termed AMY1, AMY2 and AMY3. Amylin (also known as islet amyloid polypeptide) is produced by pancreatic beta cells. Experiments with amylin-deficient mice indicate that amylin and its receptors control food intake and pain perception. Pramlintide, a synthetic analog of amylin, is used clinically for glucose control in diabetics. Chemicon's AMY1 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 AMY1 interactions with hCGRP. The membrane preparations exhibit a Kd of 0.29 nM for [125I]-hCGRP. With 10 ug/well AMY1 Membrane Prep and 0.2 nM [125I]-hCGRP, a greater than 10-fold signal-to-background ratio was obtained.
      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 10-fold signal:background with 125I-labeled hCGRP at 0.2 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 AMY1 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
      • CALCR
      • CTR
      • CTR1
      • CT-R
      • CRT
      GPCR ClassB
      Host CellsChem-1
      Protein TargetAMY1
      Target Sub-FamilyCalcitonin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P30988 # This is a receptor for calcitonin. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase. The calcitonin receptor is thought to couple to the heterotrimeric guanosine triphosphate-binding protein that is sensitive to cholera toxin.
      SIZE: 490 amino acids; 57303 Da
      SUBUNIT: Interacts with GPRASP2.
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P30988 ## Belongs to the G-protein coupled receptor 2 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: [125I]-hCGRP (Perkin Elmer # NEX354)
      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
      Product Usage Statements
      Quality AssuranceSPECIFICATIONS: 1 unit = 10 μg membrane preparation
      Bmax: 0.45 pmol/mg
      Kd: 0.29 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
      Bestellnummer GTIN
      HTS127M 04053252590115

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      Kategorien

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