由来生物
human
品質水準
リコンビナント
expressed in Chem-1 cells
形状
liquid
メーカー／製品名
ChemiScreen
Chemicon®
テクニック
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBIアクセッション番号
UniProtアクセッション番号
詳細
Full-length human CYSLTR1 cDNA
The cysteinyl leukotrienes, leukotriene C4, leukotriene D4 and 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, CysLT1 and CysLT2. A CysLT1 selective antagonist, montelukast, is used clinically in the treatment of asthma (Brink et al., 2003; Evans, 2002). Chemicon′s CysLT1 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 CysLT1 interactions 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.
アプリケーション
Human CysLT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding
生物化学的／生理学的作用
- GPCR Class: A
- Protein Target: CysLT1
- Target Sub-Family: Leukotriene
規格
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°C.
Radioligand: [3H] LTD4 (Perkin Elmer#: NET-101)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
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°C.
Radioligand: [3H] LTD4 (Perkin Elmer#: NET-101)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
物理的形状
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 with 3H-labeled LTD4 at 1.5 nM.
Liquid in packaging buffer: 50 mM
Liquid in packaging buffer: 50 mM
保管および安定性
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
法的情報
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責事項
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
保管分類コード
12 - Non Combustible Liquids
WGK
WGK 1
適用法令
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Jan Code
HTS061M:
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