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HTS116M ChemiSCREEN™ Human M3 Muscarinic Acetylcholine Receptor Membrane Preparation

Human M3 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: P20309 Entrez Gene Number: NM_000740.2
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HTS116M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1M3
      Description
      Catalogue NumberHTS116M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Human M3 Muscarinic Acetylcholine Receptor Membrane Preparation
      OverviewFull-length human CHRM3 cDNA encoding M3
      Background InformationThe muscarinic acetylcholine receptor (mAChR) family consists of five GPCRs that mediate some of the neurotransmission functions of acetylcholine in the CNS and the periphery. The M1, M3 and M5 receptors couple to Gq to mobilize intracellular calcium, whereas the M2 and M4 receptors couple to Gi/o to inhibit cAMP production (Caulfield and Birdsall, 1998). M3 is expressed prominently in smooth muscle, and plays a primary role in mediating mAChR agonist-induced contractility. Mice lacking M3 have dilated pupils, which indicates a role for M3 in regulating tone of the pupillary sphincter muscle. In addition, Msub3 plays a role in feeding, as indicated by the lean and hypophagic phenotype of M3-null mice (Wess, 2004). Chemicon's M3 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 M3 interactions with 4-DAMP. The membrane preparations exhibit a Kd of 0.72-1 nM for [3H]-4-DAMP. With 10 µg/well M3 Membrane Prep and 0.75 nM [3H]-4-DAMP, a greater than 4-fold signal-to-background ratio was 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 protein were adjusted to 0.5 mg/ml in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman M3 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 muscarinic cholinergic receptors belong to a larger family of G protein-coupled receptors. The functional diversity of these receptors is defined by the binding of acetylcholine and includes cellular responses such as adenylate cyclase inhibition, phosphoinositide degeneration, and potassium channel mediation. Muscarinic receptors influence many effects of acetylcholine in the central and peripheral nervous system. The muscarinic cholinergic receptor 3 controls smooth muscle contraction and its stimulation causes secretion of glandular tissue.
      Gene Symbol
      • CHRM3
      • HM3
      GPCR ClassA
      Host CellsChem-1
      Protein TargetM3
      Target Sub-FamilyAcetylcholine (muscarinic)
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P20309 # The muscarinic acetylcholine receptor mediates various cellular responses, including inhibition of adenylate cyclase, breakdown of phosphoinositides and modulation of potassium channels through the action of G proteins. Primary transducing effect is Pi turnover.
      SIZE: 590 amino acids; 66128 Da
      SUBCELLULAR LOCATION: Basolateral cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P20309 ## Belongs to the G-protein coupled receptor 1 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: [3H]-4-DAMP (Perkin Elmer#:NET1040 )
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM 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 4-fold signal:background with 3H labeled 4-DAMP at 0.75 nM
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information


      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of M3 membrane prep:
      Product Usage Statements
      20 µg/well 10 µg/well
      Signal:Background 5.17 5.10
      Specific Binding (cpm) 1224 1030


      SPECIFICATIONS: 1 unit = 10 µg
      Bmax: 3.75 pmol/mg
      Kd: 0.86 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
      Catalogue Number GTIN
      HTS116M 04053252678844

      Documentation

      ChemiSCREEN™ Human M3 Muscarinic Acetylcholine Receptor Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ Human M3 Muscarinic Acetylcholine Receptor Membrane Preparation Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN M3 SOMATOSTATIN RECEPTOR - 2165786 2165786
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN M3 SOMATOSTATIN RECEPTOR - 2250867 2250867

      References

      Reference overviewPub Med ID
      Muscarinic acetylcholine receptor knockout mice: novel phenotypes and clinical implications.
      Wess, Jürgen
      Annu. Rev. Pharmacol. Toxicol., 44: 423-50 (2004)  2004

      Show Abstract       		14744253 14744253
      International Union of Pharmacology. XVII. Classification of muscarinic acetylcholine receptors
      Caulfield, M P and Birdsall, N J
      Pharmacol Rev, 50:279-90 (1998)  1998

      		9647869 9647869

      Data Sheet

      Title
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN M3 SOMATOSTATIN RECEPTOR

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