Sigma-Aldrich ChemiScreen Membrane Preparation Recombinant Human TBXA2R (TP) Thromboxane Receptor，用于放射性配体结合试验和GTP γ S结合的人TP/TXA2/PGH2 GPCR膜制备。

产品概述

编码TP短型（TPα）的人全长人TBXA 2 R cDNA

TP receptors (Thromboxane A2 receptors) are widely distributed among different organ systems and have been localized on both cell membranes and intracellular structures. TP receptors belong to G protein-coupled receptor family (Hirata et al., 1991). Activation of TP receptors induces platelet aggregation, vascular and respiratory smooth muscle constriction, and enhances mitogenic responses of vascular smooth muscle cells that are stimulated by growth factors (Ali et al, 1993; Hanasaki et al., 1990). The human TP receptor is encoded by a single gene that is alternatively spliced at the carboxyl terminus, resulting in two isoforms, TPα (343 residues) and TPβ (407 residues). Both isoforms couple to Gq pathway, but couple oppositely to adenylate cyclase. The cDNA encoding human TPα has been stablely expressed in the Chem-1 host, which supports high levels of recombinant receptor expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. The membrane preparations exhibit a Kd of 9.19 nM for [3H]-SQ 29548. With 10 nM [3H]-SQ 29548, 10 ug/well TP Membrane Prep yields greater than 5-fold signal-to-background ratio.

应用领域

放射性配体结合测定和GTPγS结合。

生化/生理作用

GPCR Class：A

Protein Target：TP / TXA2 / PGH2

目标子家族：前列腺素类

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。（3 H）-SQ 29548（Perkin Elmer#NET936）洗涤缓冲液：50 mM Hepes，pH 7.4，500 mM NaCl，0.1% BSA，过滤并储存在4 ℃。

包装信息

200单位

物理形态

一个包装包含足够用于至少200次测定（单位）的膜，其中一个单位是将产生大于5倍信号的膜的量：15 nM的3 H标记的SQ 29548的背景。包装缓冲液中的液体：50 mM Tris pH 7.4，10%甘油和1% BSA，不含防腐剂。将膜蛋白在包装缓冲液中调节至指定浓度，快速冷冻，并储存在-80 ° C下。

制备/使用说明

在-70 ° C下储存。当按照指示储存时，产品自收到之日起至少6个月保持稳定。请勿冷冻和解冻。

分析说明

信号：使用不同量的Y2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5 µg/孔信号：背景6.6 4.6特异性结合（cpm）2090 963 1单位= 10 μg膜制备物Bmax：13.9 pmol/mg Kd：14.6 nM

技术规格