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HTS115M ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor

Human M2 GPCR membrane prepation for Radioligand binding Assays.

ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P08172 Entrez Gene Number: NM_000739.2, NM_001006626.1, NM_001006627.1, NM_001006628.1, NM_001006629.1, NM_001006630.1, NM_001006631.1, NM_001006632.1
HTS115M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1M2
Description
Catalogue NumberHTS115M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor
OverviewFull-length human CHRM2 cDNA encoding M2
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). In urinary bladder trachea and stomach, M2 augments the function of M3 in promoting contractility, and activation of M2 serves to counteract relaxation induced by increased cAMP levels (Ehlert et al., 2005; Wess, 2004). In addition, the ability of mAChR agonists to decrease heart rate appears to be mediated primarily by M2. Agonists of mAChRs induce tremor, hypothermia, corticosterone release, and analgesia; each of these functions is mediated at least in part by M2 (Wess, 2004). Chemicon's M2 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 M2 interactions with its ligands. The membrane preparations exhibit a Kd of 0.24 nM for [3H]-Scopolamine methyl chloride (NMS). With 10 µg/well M2 Membrane Prep and 0.5 nM [3H]-NMS, greater than 3-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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Quality LevelMQ100
Applications
ApplicationHuman M2 GPCR membrane prepation for Radioligand binding Assays.
Key Applications
  • Radioligand binding assay
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 to these receptors 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 2 is involved in mediation of bradycardia and a decrease in cardiac contractility. Multiple alternatively spliced transcript variants have been described for this gene.
Gene Symbol
  • CHRM2
  • MGC120006
  • FLJ43243
  • MGC120007
  • HM2
GPCR ClassA
Host CellsChem-1
Protein TargetM2
Target Sub-FamilyAcetylcholine (muscarinic)
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P08172 # 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 adenylate cyclase inhibition.
SIZE: 466 amino acids; 51715 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P08172 ## 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 4mM Na2HPO4, 1mM KH2PO4, pH7.4. 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 and Wash buffer: 4mM Na2HPO4, 1mM KH2PO4, pH7.4, filtered and stored at 4°C Radioligand: [3H] NMS (Perkin Elmer#:NEX-636 ) One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 3-fold signal:background with 3H-labeled NMS at 0.5 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 M2 membrane prep and unlabeled Atropine.
Product Usage Statements
10 µg/well
Signal:Background 3.2
Specific Binding (cpm) 606.8
SPECIFICATIONS: 1 unit = 10 µg
Bmax for [3H]-NMS binding: 0.5 pmol/mg protein
Kd for [3H]-NMS binding: ~0.24 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
HTS115M 04053252275913

Documentation

ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor MSDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN M2 MUSCARINIC ACETYLCHOLINE RECEPTOR - 2377138 2377138
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN M2 MUSCARINIC ACETYLCHOLINE RECEPTOR - 2278660 2278660

References

Reference overviewPub Med ID
The M2 muscarinic receptor mediates contraction through indirect mechanisms in mouse urinary bladder
Ehlert, Frederick J, et al
J Pharmacol Exp Ther, 313:368-78 (2005)  2004

15608083 15608083
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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