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HTS075M ChemiScreen™ Membrane Preparation
Recombinant Human M5 Muscarinic Acetylcholine Receptor

Human M5 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: P08912 Entrez Gene Number: NM_012125.2
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HTS075M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1M5
      Description
      Catalogue NumberHTS075M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Membrane Preparation
      Recombinant Human M5 Muscarinic Acetylcholine Receptor
      OverviewFull-length human CHRM5 cDNA, encoding M5
      Background InformationThe muscarinic acetylcholine receptor family consists of five GPCRs that mediate some of the neurotransmission functions of acetylcholine in the CNS and the periphery. The M5 receptor, along with the M1 and M3 receptors, signal through Gq/11 and subsequent release of Ca++ from the ER (Caulfield and Birdsall, 1998). Although M5 is expressed in the CNS at relatively low levels, it appears to be the only muscarinic acetylcholine receptor expressed in midbrain dopaminergic neurons, and it has been shown to mediate dopamine release from these neurons. The M5 receptor is also expressed in blood vessels in the brain and the periphery, and studies with M5 knockout mice demonstrate that M5 mediates the vasodilatory action of acetylcholine on cerebral microvessels. Further studies with M5 knockout mice also indicate a role for M5 in the rewarding effects of cocaine and morphine (Wess, 2004). Chemicon's M5 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 M5 interactions with its ligands. The membrane preparations exhibit a Kd of 0.16 nM for [3H]-Scopolamine methyl chloride (NMS). With 10 μg/well M5 Membrane Prep and 1 nM [3H]-NMS, greater than 5-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 M5 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 clinical implications of this receptor are unknown; however, stimulation of this receptor is known to increase cyclic AMP levels.
      Gene Symbol
      • CHRM5
      • MGC41838
      • HM5
      GPCR ClassA
      Host CellsChem-1
      Protein TargetM5
      Target Sub-FamilyAcetylcholine (muscarinic)
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P08912 # 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: 532 amino acids; 60074 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P08912 ## 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] NMS (Perkin Elmer#:NEX-636 )
      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 5-fold signal:background with 3H-labeled NMS at 1 nM
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceBmax: 2.49 pmol/mg; Kd: 0.16nM
      1 unit = 10 μg
      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
      HTS075M 04053252737954

      Documentation

      ChemiScreen™ Membrane Preparation
      Recombinant Human M5 Muscarinic Acetylcholine Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      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

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