Sigma-Aldrich ChemiSCREEN MC3 Membrane Preparation

产品概述

人MC 3

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.

应用领域

用于放射性配体结合测定和GTP γ S结合的人MC 3 GPCR膜制备。

放射性配体结合测定和GTP γ S结合。

生化/生理作用

GPCR类：A蛋白靶标：MC 3靶标亚家族：黑皮质素

产品特点

将膜与放射性配体和未标记的竞争剂混合在过滤之前，将GF/C 96孔滤板用0.33%聚乙烯亚胺包被30分钟，然后用50 mM HEPES（pH 7.4）洗涤，将结合反应转移至滤板，并洗涤9次（每次洗涤每孔1 mL），将平板干燥并计数。25 mM Hepes，pH 7.0，1 mM MgCl 2，1.5 mM CaCl 2 2 H2O，100 mM NaCl，0.2% BSA，蛋白酶抑制剂，过滤并在4 ℃下储存。放射性配体：（125 I）MSH（Perkin Elmer#NEX 352）清洗缓冲液：25 mM Hepes，pH 7.0，1 mM MgCl 2，1.5 mM CaCl 2 2 H 2 O，100 mM NaCl，0.2% BSA，过滤并储存在4 ℃下。一个小瓶含有足够用于至少200次测定（单位）的膜，其中单位是产生大于4倍信号的膜的量：0.25 nM 125 I标记MSH的背景

物理形态

包装缓冲液中的液体：包装方法：将膜蛋白在1 ml包装缓冲液中调节至0.5mg/ml，快速冷冻，并储存在-80 ℃。

制备/使用说明

在-70 ° C下冷冻保存长达2年。不要冷冻和解冻。

技术规格