Sigma-Aldrich ChemiSCREEN Human NK1 Tachykinin Receptor Membrane Preparation，用于放射性配体结合试验和GTP γ S结合的人NK1 GPCR膜制备。

产品概述

编码NK 1的全长人TACR 1 cDNA

The tachykinin peptide family in mammals comprises three peptides, substance P, neurokinin A and neurokinin B, which bind to the tachykinin receptor family of GPCRs, NK1, NK2 and NK3 (Severini et al., 2002). Tachykinins have prominent activity in the GI system, in which they stimulate intestinal contraction and salivation. These effects are mediated by NK1 and NK2, and an antagonist of NK1, aprepitant, is used for treatment of chemotherapy-induced emesis (Rupniak and Kramer, 1999). The NK1 tachykinin receptor is expressed in brain, and is thought to be involved in depression and nociception (Saria, 1999). Although NK1 appears to promote nociception in animal models, the significance of this activity is controversial, as antagonists of NK1 have not proven efficacious in pain relief in humans (Hill, 2000). Chemicon′s NK1 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 NK1 interactions with Substance P. The membrane preparations exhibit a Kd of 0.28-0.4 nM for [125I]-Substance P. With 10 µg/well NK1 Membrane Prep and 0.25 nM [¹²⁵I]-Substance P, a greater than 5-fold signal-to-background ratio was obtained.

应用领域

放射性配体结合试验和GTP γ S结合。

生化/生理作用

GPCR Class：A

蛋白质靶标：NK 1

靶向亚家族：粘性激肽/神经激肽

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3次结合缓冲液：50 mM Hepes，pH 7.4，5 mM MgCl 2，1 mM CaCl 2，0.2% BSA，过滤并在4 ℃下储存。（125 I）-物质P（Perkin Elmer#：NEX 190）洗涤缓冲液：50 mM Hepes，pH 7.4，500 mM NaCl，0.1% BSA，过滤并储存在4 ℃。一个包装含有足够用于至少200次测定（单位）的膜，其中单位是将产生大于5倍信号的膜的量：0.25 nM 125 I标记物质P的背景

物理形态

包装缓冲液中的液体：50 mM Tris pH 7.4，10%甘油和1% BSA，无防腐剂。包装方法：将膜蛋白用1 mL包装缓冲液调节至0.5 mg/ml，迅速冷冻，-80 ° C保存。

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。

分析说明

信号：使用不同量的NK 1膜制备物在竞争性结合试验中获得的背景和特异性结合值：10 µg/孔信号：背景7.17特异性结合（cpm）2210质量标准：1单位= 10 µg Bmax：0.38 pmol/mg Kd：0.34 nM

技术规格