ChemiSCREEN Membrane Preparation Recombinant Human mGlu₆Metabotropic Glutamate Receptor

半岛bd体育手机客户端 说明

一般描述

human GRM6 cDNA
Glutamate is a main excitatory neurotransmitter in the central nervous system, and it plays a role in learning, memory and neurotoxicity. The biological actions of glutamate are mediated by ionotropic and metabotropic glutamate receptors, which are ion channels and GPCRs respectively. Metabotropic glutamate receptors (mGluRs) are members of the class 3 G-protein coupled receptor family, which are characterized by a large extracellular domain. They are further classified into group I, II, and III mGluRs on the basis of their sequence identity, pharmacology, and signal transduction mechanism. Group I (mGlu₁and mGlu₅) couple to the phospholipase C pathway through G_αq, whereas group II (mGlu₂and mGlu₃) and group III (mGlu₄, mGlu₆, mGlu₇, and mGlu₈) negatively couple to the adenylyl cyclase pathway though G_αi(Conn and Pin, 1997). mGlu₆has a restricted localization to the dendrites of ON bipolar cells that synapse with retinal rods and cones. Targeted deletion of mGlu₆results in deficits in ON responses but unchanged OFF responses to light (Masu et al., 1995). In addition, mutations in GRM6 in humans are responsible for some forms of autosomal recessive congenital stationary night blindness (Dryja et al., 2005; Zeitz et al., 2005). Chemicon′s mGlu₆membrane 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 mGlu₆interactions with its ligands. The cell line exhibits a calcium response with EC50s of 6.2 μM, 23.4 μM, and 52.7 μM for L-AP4, (RS)PPG, and L-glutamate. The membrane preparations exhibit EC50s of of 4.0 μM, 24.1 μM, and 49.3 μM for L-AP4, (RS)PPG, and L-glutamate, respectively, in a GTPγS binding assay.

应用

Human mGlu6 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding

生化/生理作用

• GPCR Class: C
• Protein Target: mGlu6
• Target Sub-Family: Glutamate (metabotropic)

质量

EC50 in GTPγS binding assay by Glutamate: ~ 49.3 μM
EC50 in GTPγS binding assay by (RS)PPG: ~ 24.1 μM
EC50 in GTPγS binding assay by L-AP4: ~ 4.04 μM

半岛bd体育手机客户端 规格

Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl₂/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled DCG IV, (2R4R) APDC, and glutamate are added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30γC. The binding reaction is transferred to a GF/B filter plate (Millipore MAHF B1H) previously prewetted with water, and washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted.

One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific L-AP4, (RS)PPG, or glutamate -stimulated [³⁵S]-GTPγS binding.

The mGlu₆membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.

外形

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membrane protein was adjusted to 1 mg/ml in packaging buffer, rapidly frozen, and stored at -80℃.

储存及稳定性

Maintain frozen at -70℃ for up to 2 years. Do not freeze and thaw.

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

基本信息

eCl@ss

32161000

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)	ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable
NCBI登记号	NM_000843.3
UniProt登记号	O15303

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