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HTS040M ChemiSCREEN™ Membrane Preparation Recombinant Human NOP Opioid Receptor

Human NOP / ORL1 / OP4 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: P35377 Entrez Gene Number: X72304
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HTS040M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1NOP / ORL1 / OP4
      Description
      Catalogue NumberHTS040M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human NOP Opioid Receptor
      OverviewFull-length human OPRL1 cDNA encoding NOP
      Background InformationThe NOP receptor (also known as ORL1) is related to the opioid receptor family of GPCRs but does not bind to classical opioids. An endogenous ligand for NOP has been characterized and termed orphanin FQ or nociceptin (OFQ/N), which in turn does not bind to other members of the opioid receptor family. NOP is expressed widely in the CNS, and binding of OFQ/N to NOP1 appears to function in nociception, locomotor activity, anxiety, reward, memory and tolerance to classical opioids (Mogil and Pasternak, 2001). Millipore's NOP 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 NOP. The membrane preparations exhibit a Kd of 0.06 nM for [125II]-Nociceptin. With 0.1 nM [125I]-Nociceptin, 5μg/well NOP Membrane Prep typically yields greater than 10-fold signal-to-background ratio.
      References
      Product Information
      FormatMembranes
      PresentationLiquid 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 1 ml packaging buffer, rapidly frozen, and stored at -80°C
      Quality LevelMQ100
      Applications
      ApplicationHuman NOP / ORL1 / OP4 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
      • OPRL1
      • OOR
      • ORL1
      GPCR ClassA
      Host CellsChem-1
      Protein TargetNOP / ORL1 / OP4
      Target Sub-FamilyOpioid
      UniProt Number
      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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50 mM Tris-HCl, 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-HCl, pH 7.4 filtered and stored at 4°C

      Radioligand: [125I]-Nociceptin. (Perkin Elmer#:NEX-324 )

      Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM 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 10-fold signal:background with 125I labeled Nociceptin
      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 NOP Receptor membrane prep.
      Product Usage Statements
      5 µg/well
      Signal:Background 23.8
      Specific Binding (cpm) 6656


      SPECIFICATIONS: 1 unit = 5 µg
      Bmax for [125I]-Nociceptin binding: 1.1 pmol/mg protein
      Kd for [125I]-Nociceptin binding: ~0.06 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 up to the expiration date indicated on the product label. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      HTS040M 04053252271250

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