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Figure 1. Saturation binding for M2. 10 µg/well M2 Membrane Preparation was incubated with increasing amount of 3. ...
Figure 2. Competition binding for M2. 10 µg/well M2 Membrane Preparation (HTS115M) was incubated with 0.5 nM 3H-lab...
ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor
Description:
ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
200 units
Product Overview:
Full-length human CHRM2 cDNA encoding M2
Background Information:
The 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.
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.
Entrez Gene Summary:
The 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.
UniProt Summary:
FUNCTION: 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.
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.
Species:
Human
Brand Family:
Chemicon
Protein Target:
M2
Presentation:
Liquid 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.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Ligand Type:
non-peptide
Target Sub-Family:
Acetylcholine (muscarinic)
Storage Conditions:
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
GPCR Class:
A
Promotional Text:
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Gene Symbol:
- CHRM2
- MGC120006
- FLJ43243
- MGC120007
- HM2
Protein or Enzyme Type:
GPCR Membrane Preparations
Product Name:
ChemiSCREEN™ Membrane Preparation Recombinant Human M2 Muscarinic Acetylcholine Receptor
Alternate Names:
mAChR2
Quality Assurance:
Table 1. Signal:background and specific binding values obtained in a competition binding assay with M2 membrane prep and unlabeled Atropine.
SPECIFICATIONS: 1 unit = 10 µg
Bmax for [3H]-NMS binding: 0.5 pmol/mg protein
Kd for [3H]-NMS binding: ~0.24 nM
| 10 µg/well | |
|---|---|
| Signal:Background | 3.2 |
| Specific Binding (cpm) | 606.8 |
Bmax for [3H]-NMS binding: 0.5 pmol/mg protein
Kd for [3H]-NMS binding: ~0.24 nM
Cell Type:
Chem-1
Incubation 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 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
Host Cells:
Chem-1
UniProt Number:
Entrez Gene Number:
Cell Membrane Type:
GPCR