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HTS019M ChemiSCREEN™ CB1 Membrane Preparation

Human CB1 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: P21554 Entrez Gene Number: NM_016083.3, NM_033181.2
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HTS019M
200 assays  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1CB1
      Description
      Catalogue NumberHTS019M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ CB1 Membrane Preparation
      Overview Human CB1
      Background InformationCB1 is a GPCR that is expressed primarily in brain and nervous tissue, and mediates numerous CNS responses such as analgesia, appetite, cognition, memory and locomotor activity. A number of cannabinoid ligands bind to CB1 and activate Gi/o-mediated downstream responses, including inhibition of cAMP production and activation of ion channels and MAP kinases. Ligands for CB1 include exogenous agonists such as Delta9-THC, the main psychoactive component of the plant Cannabis sativa, and endogenous eicosanoid agonists such as anandamide. A number of synthetic agonists such as CP55940 and R-(+)-WIN55212, and antagonists, such as SR141716A, for CB1 have been developed (Howlett et al., 2002). CB1 agonists have clinical utility in analgesia and antiemetic properties, whereas CB1 antagonists show promise for treatment of appetite in obesity disorders. Chemicon's CB1 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 for agonists and antagonists at CB1. The membrane preparations exhibit a Kd of 7 nM for [3H]-SR141716A. In the presence of 2 nM [3H]-SR141716A with CP55940 as unlabeled competitor, 5 μg/well and 10 μg/well CB1 Membrane Prep yield 4- and 5-fold signal-to-background ratios, respectively.
      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.
      Quality LevelMQ100
      Applications
      ApplicationHuman CB1 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
      Concentration1.0 mg/mL
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a protein that is one of two cannabinoid receptors. The cannabinoids, principally delta-9-tetrahydrocannabinol and synthetic analogs, are psychoactive ingredients of marijuana. The cannabinoid receptors are members of the family of guanine-nucleotide-binding protein (G-protein) coupled receptors which inhibit adenylate cyclase activity in a dose-dependent, stereoselective and pertussis toxin-sensitive manner. The two receptors have been found to be involved in the cannabinoid-induced CNS effects (including alterations in mood and cognition) experienced by users of marijuana. Two transcript variants encoding different isoforms have been described for this gene.
      Gene Symbol
      • CNR1
      • CNR
      • CANN6
      • CB1A
      • CB1
      • CB1K5
      • CB-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetCB1
      Target Sub-FamilyCannabinoid
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P21554 # Involved in cannabinoid-induced CNS effects. Acts by inhibiting adenylate cyclase. Could be a receptor for anandamide. Isoform 2 and isoform 3 have altered ligand binding.
      SIZE: 472 amino acids; 52858 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Widely expressed.
      SIMILARITY: SwissProt: P21554 ## 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, 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. Ligands were diluted in binding buffer containing 30% DMSO, then added to membranes such that the final DMSO concentration was 15%.Radioligand: [3H] SR141716A (Amersham #TRK1028)

      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 approximately 4-fold signal:background with [3H] SR141716A at 2 nM
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceSPECIFICATIONS:
      1 unit = 10 μg
      Bmax: 15 pmol/mg
      Kd: 7 nM for SR141716A
      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 assays
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      HTS019M 04053252624063

      Documentation

      Protocols

      Title
      Assembly of a Competition Binding Curve

      ChemiSCREEN™ CB1 Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ CB1 Membrane Preparation Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CB1 CANNABINOID RECEPTOR - 2341139 2341139
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CB1 CANNABINOID RECEPTOR - 2235014 2235014

      Data Sheet

      Title
      CHEMISCREEN¿ MEMBRANE PREPARATION RECOMBINANT HUMAN CB%26lt;sub%26gt;1%26lt;/sub%26gt; CANNABINOID RECEPTOR - Data Sheet

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