Sigma-Aldrich ChemiScreen VPAC1 Membrane Preparation，用于放射性配体结合测定和GTP γ S结合的人VPAC 1/VIP 1 GPCR膜制备。

产品概述

人VIPR 1 cDNA，编码VPAC 1

Vasoactive intestinal peptide (VIP), a 28 amino acid peptide originally isolated by its vasodilation activity, binds to two class B GPCRs, VPAC1 and VPAC2, to exert its functions in the CNS, vasculature, immune system and adrenal medulla (Harmar et al., 1998). In the immune system, VIP is synthesized by mast cells and lymphocytes, and appears to inhibit inflammation and to shift the immune response toward a Th2 pathway (Delgado et al., 2004). In the heart, VIP is expressed by nerve fibers, where it modulates heart rate, and coronary blood flow (Henning and Sawmiller, 2001). Chemicon′s VPAC1 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 VPAC1 interactions with VIP. The membrane preparations exhibit a Kd of 1.2 nM for [125I]-VIP. With 2.5 μg/well VPAC1 Membrane Prep and 0.4 nM [125I]-VIP, a greater than 5-fold signal-to-background ratio is obtained.

应用领域

放射性配体结合测定和GTP γ S结合。

生化/生理作用

GPCR类别：B

Protein Target：VPAC1 /VIP 1

目标子系列：VIP/PACAP

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 HEPES, pH 7.4, 0.5% BSA. 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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C

Radioligand: [125I] VIP (Perkin Elmer# NEX192)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 5-fold signal:background with 125I-labeled VIP at 0.4 nM

物理形态

包装缓冲液中的液体：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 ° C。

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：在竞争结合试验中使用不同量的VPAC 1膜制备物获得的背景和特异性结合值：10 μg/孔5 μg/孔信号：背景9.7 10.8特异性结合（cpm）30299 31224规格：1单位= 5 μg Bmax：19.65 pmol/mg Kd：1.2 nM

技术规格