ChemiSCREEN Membrane Preparation Recombinant Human CysLT₁Cysteinyl Leukotriene Receptor

半岛bd体育手机客户端 说明

一般描述

The cysteinyl leukotrienes, leukotriene C₄, leukotriene D₄and leukotriene E₄, are arachidonic acid derivatives modified by glutathione, Cys-Gly or Cys. Activated mast cells release cysteinyl leukotrienes, which cause smooth muscle contraction, airway constriction, and vascular permeability. The biological effects of the cysteinyl leukotrienes are mediated by two GPCRs, CysLT₁and CysLT₂. A CysLT₁selective antagonist, montelukast, is used clinically in the treatment of asthma (Brink et al., 2003; Evans, 2002). Chemicon′s CysLT₁membrane 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 CysLT₁interactions and its ligands. The membrane preparations exhibit a Kd of 0.97nM for [³3H]-Leukotriene D₄(LTD4). With 10.0ug/well CysLT1 membrane prep and 1.5 nM [³3H]-Leukotriene D₄, a greater than 3-fold signal-to-background ratio was obtained.
Full-length human CYSLTR1 cDNA

应用

Human CysLT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding

生化/生理作用

• GPCR Class: A
• Protein Target: CysLT1
• Target Sub-Family: Leukotriene

半岛bd体育手机客户端 规格

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 50 mM 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, pH7.4, 20 mM CaCl₂, 25 mM MgCl₂, 10 mM Glycine, 10 mM L-Cysteine, filtered and stored at 4℃.

Radioligand: [³H] LTD4 (Perkin Elmer#: NET-101)

Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4℃.

外形

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 3-fold signal:background with³H-labeled LTD4 at 1.5 nM.

Liquid in packaging buffer: 50 mM

储存及稳定性

Maintain frozen at -70℃ for up to 2 years. Do not freeze and thaw.

法律信息

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

免责声明

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial

基本信息

eCl@ss

32161000

半岛bd体育手机客户端 性质

质量水平	100
生物来源	human
重组	expressed in Chem-1 cells
manufacturer/tradename	ChemiScreen Chemicon?
technique(s)	ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable
NCBI登记号	NM_006639.2
UniProt登记号	Q9Y271

安全信息

储存分类代码	12 - Non Combustible Liquids
WGK	WGK 1
闪点(F)	Not applicable
闪点(C)	Not applicable