Sigma-Aldrich ChemiSCREEN Human δ Opioid Receptor Membrane Preparation，人Delta/OP 1/DOP/DOR GPCR膜的制备及GTP γ S的放射性配体结合分析。

产品概述

编码δ阿片受体的全长人OPRD 1 cDNA

Opiates derived from the opium poppy, Papaver somniferum, have been used in for millenia for their anti-diarrheal, analgesic and euphoric properties. More recently, endogenous peptides, enkephalins, dynorphins, and endorphins, were found to bind to the same sites as opiate alkaloids. The receptors for the classical opioids are three related GPCRs, δ, κ, and μ (also known as OP1, OP2 and OP3, resepctively), that activate Gi/o to reduce intracellular cAMP levels. Although most clinically used opioids function by activation of the µ opioid receptor, agonits of spinal δ opioid receptors have antinociceptive activity that is independent of µ. In addition, activation of δ increases locomotor activity, inhibits gastrointestinal motility, and decreases respiratory frequency (Dhawan et al., 1996). Agonists for δ opioid receptors also exhibit antidepressant-like activity in animal models (Broom et al., 2002). Chemicon′s δ 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 agonists and antagonists at δ. The membrane preparations exhibit a Kd of 1.12 nM for [3H]-DADLE. With 0.9 nM [3H]-DADLE, 10 µg/well δ Membrane Prep typically yields greater than 4-fold signal-to-background ratio.

应用领域

放射性配体结合试验和GTPγS

生化/生理作用

GPCR Class：A

Protein Target：Delta / OP1 / DOP / DOR

目标子家族：阿片类

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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM Tris-HCl. 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-HCl, filtered and stored at 4°C
Radioligand: [3H] DADLE (Perkin Elmer#: NET-648)
Wash Buffer: 50 mM Tris-HCl, 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：具有0.9 nM 3 H标记的DADLE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lmg/mL，并以lmL/小瓶分配。

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

Signal:background and specific binding values obtained in a competition binding assay with δ membrane prep and unlabeled DPDPE.
10 g/well
Signal:Background: 7.32
Specific Binding (cpm): 955

SPECIFICATIONS: 1 unit = 10 g
Bmax for [3H] DADLE binding: 11.6 pmol/mg protein
Kd for [3H] DADLE binding: ~1.12 nM

技术规格