由来生物
human
品質水準
リコンビナント
expressed in Chem-1 cells
形状
liquid
メーカー／製品名
ChemiScreen
Chemicon®
テクニック
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBIアクセッション番号
UniProtアクセッション番号
詳細
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 (mGlu1 and mGlu5) couple to the phospholipase C pathway through Gαq, whereas group II (mGlu2 and mGlu3) and group III (mGlu4, mGlu6, mGlu7, and mGlu8) negatively couple to the adenylyl cyclase pathway though Gαi (Conn and Pin, 1997). mGlu6 has a restricted localization to the dendrites of ON bipolar cells that synapse with retinal rods and cones. Targeted deletion of mGlu6 results 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 mGlu6 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 mGlu6 interactions 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.
human GRM6 cDNA
アプリケーション
Human mGlu6 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding
生物化学的／生理学的作用
- GPCR Class: C
- Protein Target: mGlu6
- Target Sub-Family: Glutamate (metabotropic)
品質
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
EC50 in GTPγS binding assay by (RS)PPG: ~ 24.1 μM
EC50 in GTPγS binding assay by L-AP4: ~ 4.04 μM
規格
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 mGlu6 membrane 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.
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 mGlu6 membrane 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.
物理的形状
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°C.
Packaging method: Membrane protein was adjusted to 1 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C.
保管および安定性
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
法的情報
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責事項
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
保管分類コード
12 - Non Combustible Liquids
WGK
WGK 2
適用法令
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Jan Code
HTS149M:
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