ChemiSCREEN Membrane Preparation Recombinant Human M₂Muscarinic Acetylcholine Receptor

半岛bd体育手机客户端 说明

一般描述

Full-length human CHRM2 cDNA encoding M₂
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, M₃and M₅receptors couple to G_qto mobilize intracellular calcium, whereas the M₂and M₄receptors couple to G_i/oto inhibit cAMP production (Caulfield and Birdsall, 1998). In urinary bladder trachea and stomach, M₂augments the function of M₃in promoting contractility, and activation of M₂serves 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 M₂membrane 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 M₂interactions with its ligands. The membrane preparations exhibit a Kd of 0.24 nM for [³H]-Scopolamine methyl chloride (NMS). With 10 ?g/well M₂Membrane Prep and 0.5 nM [³H]-NMS, greater than 3-fold signal-to-background ratio was obtained.

生化/生理作用

Protein Target: M2
GPCR Class: A
Target Sub-Family: Acetylcholine (muscarinic)

质量

Table 1. Signal:background and specific binding values obtained in a competition binding assay with M₂membrane prep and unlabeled Atropine.

	10 ?g/well
Signal:Background	3.2
Specific Binding (cpm)	606.8

SPECIFICATIONS: 1 unit = 10 ?g
Bmax for [ ³H]-NMS binding: 0.5 pmol/mg protein
K _dfor [ ³H]-NMS binding: ~0.24 nM

半岛bd体育手机客户端 规格

Inucbation 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 with³H-labeled NMS at 0.5 nM

外形

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.

法律信息

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

免责声明

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.

基本信息

eCl@ss	32161000
NACRES	NA.41

半岛bd体育手机客户端 性质

质量水平	100
生物来源	human
重组	expressed in Chem-1 cells
manufacturer/tradename	ChemiScreen Chemicon?
technique(s)	radioligand binding assay (RLBA): suitable
NCBI登记号	NM_000739.2 NM_001006626.1 NM_001006627.1 NM_001006628.1 NM_001006629.1 NM_001006630.1 NM_001006631.1 NM_001006632.1
UniProt登记号	P08172
运输	dry ice
Gene Information	human ... CHRM2(1129)

安全信息

储存分类代码	12 - Non Combustible Liquids
WGK	WGK 2
闪点(F)	Not applicable
闪点(C)	Not applicable