Sigma-Aldrich ChemiScreen Bradykinin B2 Membrane Preparation，用于放射性配体结合测定和GTPgammaS结合的人B2 GPCR膜制备。

产品概述

Bradykinins bind and activate two GPCRs, termed B1 and B2 (gene names BDKRB1 and BDKRB2, respectively), which signal through Gq and Gi. B2 is widely expressed in a constitutive fashion, whereas B1 expression is induced during inflammation. B2 appears to mediate much of the activity of kinins in vasodilation, arthritis, edema, and nociception (Leeb-Lundberg et al., 2005). Chemicon′s B2 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 B2 interactions with bradykinin. The membrane preparations exhibit a Kd of 0.18 nM for [3H]-bradykinin. With 5.0 μg/well, B2 Membrane Prep and 0.4 nM [3H]-bradykinin, a greater than 4-fold signal-to-background ratio is obtained.

全长人BDKRB 2

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：B2

目标子家族：缓激肽

产品特点

Inucbation Conditions
RECOMMENDED ASSAY 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: [3H] Bradykinin, [2,3-prolyl-3,4-3H(N)] (Perkin Elmer #NET-706)

Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM 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 4-fold signal:background with 3H-labeled bradykinin 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，并在-80 ° C下储存。

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

质量标准：1单位= 10 µg Bmax：2.73 pmol/mg Kd：0.18 nM

技术规格