Sigma-Aldrich ChemiSCREEN Human D3 Dopamine Receptor Membrane Preparation，用于放射性配体结合试验和GTPγS结合的人D3 GPCR膜制备。

产品概述

Dopamine is a catecholamine neurotransmitter that functions in the CNS to control locomotor, cognitive, emotional and neurendocrine processes, and in the periphery to modulate cardiovascular, renal and gastrointestinal processes. The biological activities of dopamine are mediated by a family of five GPCRs. The D1 and D5 subtypes couple to Gs to increase intracellular cAMP, whereas the D2, D3 and D4 subtypes couple to Gi to reduce cAMP (Missale et al., 1998). The dopamine D3 receptor is expressed in phylogenetically older regions of the brain, suggesting that it plays a role in cognitive and emotional functions. It has been the target of potential drugs for psychotic disorders and Parkinson′s disease. Chemicon′s D3 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 D3 interactions and its ligands. The membrane preparations exhibit a Kd of 0.1 nM for [125I]-Iodospiperone. With 5.0ug/well D3 membrane prep and 0.1 nM [125I]-Iodospiperone, a greater than 4-fold signal-to-background ratio was obtained.

人D3受体

应用领域

用于放射性配体结合试验和GTPγS结合的人D3 GPCR膜制备。

生化/生理作用

蛋白质靶标：D3

产品特点

Inucbation Conditions
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 Tris, pH 7.4. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Tris, pH 7.4, 120mM NaCl, filtered and stored at 4°C
Radioligand: [125I] -Iodospiperone(Perkin Elmer # NEX284)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.

物理形态

一个包装包含足够用于至少200次测定（单位）的膜，其中单位是将产生大于4倍信号的膜的量：背景为0.1 nM的125 I标记的Iodospiperone。包装缓冲液中的液体：50 mM Tris，pH 7.4，10%甘油，不含防腐剂。包装方法：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 ℃。

分析说明

表1.信号：使用不同量的D3受体膜制备物10 µg/孔5 µg/孔在竞争性结合试验中获得的背景和特异性结合值信号：背景4.88 5.93特异性结合（cpm）1853.7 1861.3规格：1单位= 5 µg膜制备物Bmax：0.32 pmol/mg Kd：0.1 nM

技术规格