半岛bd体育手机客户端 说明
一般描述
The endothelin family of peptides are potent vasoconstrictors synthesized by endothelial cells in both constitutive and inducible pathways. The biological actions of endothelins are mediated by two GPCRs, ETAand ETB(Davenport, 2002). Whereas ETAis the predominant receptor on smooth muscle cells and thus is the primary mediator of the vasoconstrictor activity, ETBis found on endothelial cells and mediates vasodilation (Nilsson et al., 1997). In addition, ETBhas also been linked to renal failure, congestive heart failure, atherosclerosis, and pulmonary hypertension (D’Orléans-Juste, et al., 2002). A mutation in ETBhas been found to cause Hirschsprung disease-2, a degenerative disease of the digestive tract (Puffenberger et al, 1994). Millipore’s cloned human ETBexpressing cell line is made in the CHO host, which supports high levels of recombinant ETBexpression on the cell surface. Thus, the ETBMembrane Preparation is an ideal tool for screening for compounds binding to ETB. The membrane preparations exhibit a Kd of 0.03-0.06 nM for [125I]-endothelin-1. With 0.2 nM [125I]-endothelin-1, 10 μg/well ETBMembrane Prep yields greater than 5 fold signal-to-background ratio.
Human ETBisoform 1
应用
Radioligand binding assay and GTPγS binding
生化/生理作用
Protein Target: ETB
GPCR Class: A
Target Sub-Family: Endothelin
质量
Table 1. Signal:background and specific binding values obtained in a competition binding assay with ETBmembrane prep.
| 10 ?g/well | |
|---|---|
| Signal:Background | 7.4 |
| Specific Binding (cpm) | 5704 |
半岛bd体育手机客户端 规格
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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) 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]Endothelin-1 (Perkin Elmer # NEX259)
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 an unit is the amount of membrane that will yield greater than 5-fold signal:background with125I-labeled Endothelin-1 at 0.25 nM.
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 5-fold signal:background with125I-labeled Endothelin-1 at 0.25 nM.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
储存及稳定性
Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. 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 uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
基本信息
| eCl@ss | 32161000 |
| NACRES | NA.84 |
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 CHO cells |
| manufacturer/tradename | ChemiScreen Chemicon? |
| technique(s) | ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable |
| NCBI登记号 | NM_00015 |
| UniProt登记号 | P24530 |
| 运输 | dry ice |
安全信息
| 储存分类代码 | 12 - Non Combustible Liquids |
| WGK | WGK 2 |
| 闪点(F) | Not applicable |
| 闪点(C) | Not applicable |
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