Sigma-Aldrich ChemiScreen BLT1 Membrane Preparation，用于放射性配体结合测定和GTP γ S结合的人BLT 1 GPCR膜制备。

产品概述

BLT1 receptor is a Gi-coupled GPCR that binds primarily to the eicosanoid leukotriene B4. Macrophages, leukocytes and eosinophils express BLT1, and in vivo, BLT1 appears to mediate leukocyte migration and inflammation (reviewed in Brink et al., 2003). Chemicon′s BLT1 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 BLT1 interactions with Leukotriene B4. The membrane preparations exhibit a Kd of 1.3 nM for [3H]-Leukotriene B4. With 5.0 μg/well BLT1 Membrane Prep and 1 nM [3H]-Leukotriene B4, a signal-to-background ratio of approximately 3-fold is obtained.

人BLT 1

应用领域

用于放射性配体结合测定和GTP γ S结合的人BLT 1 GPCR膜制备。

放射性配体结合测定和GTP γ S结合。

生化/生理作用

GPCR Class：A

蛋白质靶标：BLT 1

靶标亚家族：白三烯

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] Luekotriene (Perkin Elmer# NEX378)

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 2.5-fold signal:background or 500 cpm with 3H-labeled Leukotriene B4 at 1 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

规格说明：信号：在竞争结合试验中用不同量的胃动素受体膜制备物获得的背景和特异性结合值：5 μg/孔10 μg/孔信号：背景75 65特异性结合（cpm）46348 54457 1单位= 5μg膜制备物Bmax：5. 6 pmol/mg Kd：0. 43 nM

技术规格