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HTS172M ChemiScreen™Human CGRP1 Calcitonin Receptor Membrane Preparation

Human CGRP1 GPCR membrane prepation for Radioligand binding Assays.

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Host Cell: Chem-1 Uni Prot Number: Q16602 Entrez Gene Number: NM_005795.4
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HTS172M
200 units  
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      Replacement Information

      Tableau de caractéristiques principal

      SpeciesHost CellsProtein Target
      Human Chem-1CGRP1
      Description
      Catalogue NumberHTS172M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™Human CGRP1 Calcitonin Receptor Membrane Preparation
      OverviewFull-length human CALCRL and RAMP1 cDNAs encoding CGRP1
      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, CT receptor (calcitonin receptor) and CL receptor (calcitonin receptor-like receptor) (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. CGRP (calcitonin gene-related peptide) induces vasodilation, and its activity is primarily mediated by CGRP1, composed of CL in complex with RAMP1 (Hasbak et al., 2003). As a consequence of its role as a vasodilator, CGRP acting via CGRP1 appears to play an important role in hypertension (Deng and Li, 2005). Millipore's CGRP1 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 CGRP1 interactions with CGRP. The membrane preparations exhibit a Kd of 1.1-1.7 nM for [125I]-hCGRP. With 5 µg/well CGRP1 Membrane Prep and 1 nM [125I]-hCGRP, a greater than 15-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 15-fold signal:background with 125I-labeled hCGRP at 1 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 CGRP1 GPCR membrane prepation for Radioligand binding Assays.
      Key Applications
      • Radioligand binding assay
      Biological Information
      Entrez Gene Number
      Gene Symbol
      • CALCRL
      • CGRPR1-PEN
      • CGRPR1
      • CGRPR
      • CRLR
      GPCR ClassB
      Host CellsChem-1
      Protein TargetCGRP1
      Target Sub-FamilyCalcitonin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q16602 # Receptor for calcitonin-gene-related peptide (CGRP) together with RAMP1 and receptor for adrenomedullin together with RAMP2 or RAMP3 (By similarity). The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
      SIZE: 461 amino acids; 52979 Da
      SUBUNIT: Heterodimer of CALCRL and RAMP1, RAMP2 or RAMP3 (By similarity).
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Predominantly expressed in the lung and heart.
      SIMILARITY: SwissProt: Q16602 ## 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 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] hCGRP (Perkin Elmer # NEX354)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      SPECIFICATIONS:
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CGRP1 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 26.5 29.3
      Specific Binding (cpm) 55556 47919


      1 unit = 5 µg membrane preparation
      Bmax: 10.8 pmol/mg
      Kd: 1.4 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 Size200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Référence GTIN
      HTS172M 04053252583933

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      Catégories

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