跳转至内容
Merck
所有图片(1)

Key Documents

HTS022M

Sigma-Aldrich

ChemiSCREEN MC3 Membrane Preparation

登录查看公司和协议定价


About This Item

分類程式碼代碼:
41106514
eCl@ss:
32161000
NACRES:
NA.45

生物源

human

品質等級

重組細胞

expressed in Chem-1 cells

形狀

liquid

製造商/商標名

ChemiScreen
Chemicon®

技術

ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable

NCBI登錄號

UniProt登錄號

一般說明

Human MC3
The melanocortins, the peptides alpha-, beta- and gamma-melanocyte-stimulating hormone (MSH), bind to a family of five Gs-coupled GPCR receptors (MCRs) and play important roles in energy balance, reproductive function, pigmentation and inflammation (Gantz and Fong, 2003). MC3R is expressed in the CNS, including the hypothalamus, and appears to play a role in energy homeostasis as indicated by the observation that MC3R-null mice have increased subcutaneous fat, a loss of lean body mass, and reduced energy expenditure (Butler et al., 2000; Chen et al., 2000). Macrophages also express MC3R, which together with MC1R mediates the anti-inflammatory activity of melanocortins in animal models of peritonitis and myocardial infarct (Getting et al., 2003; 2004). Chemicon′s MC3R 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 antagonists of MC3R interactions with melanocortins. The membrane preparations exhibit a Kd of 0.4 nM for [125I]-MSH. With 10.0 mg/well MC3R Membrane Prep and 0.25 nM [125I]-MSH, a greater than 7-fold signal-to-background ratio is obtained.

應用

Human MC3 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Radioligand binding assay, and GTPgammaS binding.

生化/生理作用

  • GPCR Class: A
  • Protein Target: MC3
  • Target Sub-Family: Melanocortin

規格

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, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 9 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.Binding buffer: 25 mM Hepes, pH 7.0, 1 mM MgCl2, 1.5 mM CaCl2 2H2O, 100mM NaCl, 0.2% BSA, Protease Inhibitor, filter and stored at 4 °C.

Radioligand: [125I] MSH (Perkin Elmer# NEX352)

Wash Buffer: 25 mM Hepes, pH 7.0, 1 mM MgCl2, 1.5 mM CaCl2 2H2O, 100mM NaCl, 0.2% BSA, filtered and stored at 4°C. One vial contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 4-fold signal:background with 125I-labeled MSH at 0.25 nM

外觀

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.

儲存和穩定性

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

法律資訊

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

免責聲明

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

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類(WGK)

WGK 2


分析证书(COA)

输入产品批号来搜索 分析证书(COA) 。批号可以在产品标签上"批“ (Lot或Batch)字后找到。

已有该产品?

在文件库中查找您最近购买产品的文档。

访问文档库

我们的科学家团队拥有各种研究领域经验,包括生命科学、材料科学、化学合成、色谱、分析及许多其他领域.

联系技术服务部门