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ChemiSCREEN Membrane Preparation Recombinant Human ETB Endothelin Receptor

品牌
Sigma-Aldrich
货号
HTS046M
规格纯度
Human ETB GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
参考价格
10463.8 *本价格含增值税费
促销
数量
-+
产品介绍:

产品说明

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, ETA and ETB (Davenport, 2002). Whereas ETA is the predominant receptor on smooth muscle cells and thus is the primary mediator of the vasoconstrictor activity, ETB is found on endothelial cells and mediates vasodilation (Nilsson et al., 1997). In addition, ETB has also been linked to renal failure, congestive heart failure, atherosclerosis, and pulmonary hypertension (D’Orléans-Juste, et al., 2002). A mutation in ETB has been found to cause Hirschsprung disease-2, a degenerative disease of the digestive tract (Puffenberger et al, 1994). Millipore’s cloned human ETB expressing cell line is made in the CHO host, which supports high levels of recombinant ETB expression on the cell surface. Thus, the ETB Membrane 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 ETB Membrane Prep yields greater than 5 fold signal-to-background ratio.
Human ETB isoform 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 ETB membrane prep.

10 µg/well
Signal:Background 7.4
Specific Binding (cpm) 5704


SPECIFICATIONS: 1 unit = 10 μg membrane preparation
Bmax: 0.49 pmol/mg
Kd: 0.03-0.06 nM

产品规格

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 with 125I-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 with 125I-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@ss32161000
NACRESNA.84

产品性质

质量水平100
生物来源human
重组expressed in CHO cells
manufacturer/tradenameChemiScreen
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
WGKWGK 2
闪点(F)Not applicable
闪点(C)Not applicable

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