HTS020M ChemiSCREEN™ Membrane Preparation Recombinant Human CB2 Cannabinoid Receptor

HTS020M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human CHO (Chinese hamster ovary cells) CB2
      Description
      Catalogue NumberHTS020M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human CB2 Cannabinoid Receptor
      OverviewFull-length Human CB2
      Background InformationCannabinoid compounds include exogenous drugs such as D9-THC, the main psychoactive component of the plant Cannabis sativa, and endogenous mediators, such as anandamide, that belong to eicosanoid family.The biological effects of cannabinoids are mediated by a family of two Gi-coupled 7-transmembrane receptors, CB1 and CB2. The CB1 receptor is found primarily in brain and mediates the psychoactive effects of cannabinoid ligands. The CB2 receptor is expressed mainly in immune cells, including mast cells and CD40-activated B cells, where it mediates proliferation and inhibition of migration (Howlett et al., 2002). Activation of CB2 inhibits the development of liver fibrosis (Julien et al., 2005). In bone, CB2 is expressed in both osteoblasts and osteoclasts, and functions to prevent bone loss (Ofek et al., 2006). In addition, activation of CB2 has an antinociceptive effect in animal models of neuropathic, inflammatory, and acute pain; this effect is mediated by release of endogenous opioids in the periphery (Ibrahim et al., 2005). Millipore's CB2 membrane preparations are crude membrane preparations made from stable recombinant cell lines with a high-level of GPCR surface expression; thus, they are ideal HTS tools for screening for agonists and antagonists of CB2. The membrane preparations exhibit a Kd of 2.7 nM for [3H]-CP55,940. With 5 μg/well CB2 Membrane Prep and 2.5 nM [3H]-CP55,940, greater than 10-fold signal-to-background ratio is obtained.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
      Packaging method: Membranes proteins were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Applications
      ApplicationHuman CB2 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
      Entrez Gene SummaryThe cannabinoid delta-9-tetrahydrocannabinol is the principal psychoactive ingredient of marijuana. The proteins encoded by this gene and the cannabinoid receptor 1 (brain) (CNR1) gene have the characteristics of a guanine nucleotide-binding protein (G-protein)-coupled receptor for cannabinoids. They inhibit adenylate cyclase activity in a dose-dependent, stereoselective, and pertussis toxin-sensitive manner. These proteins have been found to be involved in the cannabinoid-induced CNS effects (including alterations in mood and cognition) experienced by users of marijuana. The cannabinoid receptors are members of family 1 of the G-protein-coupled receptors.
      Gene Symbol
      • CNR2
      • CB-2
      • OTTHUMP00000044841
      • CB2
      • CX5
      GPCR ClassA
      Host CellsCHO (Chinese hamster ovary cells)
      Protein TargetCB2
      Target Sub-FamilyCannabinoid
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P34972 # Involved in cannabinoid-induced CNS effects through G- protein mediated inhibition of adenylate cyclase. Could be a receptor for anandamide.
      SIZE: 360 amino acids; 39681 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Not found in brain. Found in spleen.
      SIMILARITY: SwissProt: P34972 ## Belongs to the G-protein coupled receptor 1 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) 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: [3H]-CP55940 (Perkin Elmer #NET1051)

      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 an unit is the amount of membrane that will yield greater than 10-fold signal:background with 3H-labeled CP55,940 at 2.5 nM
      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 varying amounts of CB2 membrane prep.
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 33.5 25.2
      Specific Binding (cpm) 7336 6207

      SPECIFICATIONS: 1 unit = 5 μg membrane preparation
      Bmax: 50 pmol/mg
      Kd: 2.7 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

      Documentation

      ChemiSCREEN™ Membrane Preparation Recombinant Human CB2 Cannabinoid Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Peripheral cannabinoid receptor, CB2, regulates bone mass.
      Ofek, Orr, et al.
      Proc. Natl. Acad. Sci. U.S.A., 103: 696-701 (2006)  2006

      Show Abstract
      16407142 16407142

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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