ChemiScreen™ Membrane Preparation Human Recombinant ETB

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ChemiScreen™ Membrane Preparation Human Recombinant ETB

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货号：HTS046M

品牌：Millipore 密理博

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北京朝阳

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EA (预计3-5工作日到货)

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Replacement Information

重要规格表

Species	Host Cells	Protein Target
Human	CHO (Chinese hamster ovary cells)	ETB

Description
Catalogue Number	HTS046M
Brand Family	Chemicon®
Trade Name	ChemiScreen Chemicon
Description	ChemiSCREEN™ Membrane Preparation Recombinant Human ET_B Endothelin Receptor
Overview	Human ET_B isoform 1
Background Information	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, ET_A and ET_B (Davenport, 2002). Whereas ET_A is the predominant receptor on smooth muscle cells and thus is the primary mediator of the vasoconstrictor activity, ET_B is found on endothelial cells and mediates vasodilation (Nilsson et al., 1997). In addition, ET_B has also been linked to renal failure, congestive heart failure, atherosclerosis, and pulmonary hypertension (D’Orléans-Juste, et al., 2002). A mutation in ET_B has been found to cause Hirschsprung disease-2, a degenerative disease of the digestive tract (Puffenberger et al, 1994). Millipore’s cloned human ET_Bexpressing cell line is made in the CHO host, which supports high levels of recombinant ET_B expression on the cell surface. Thus, the ET_B Membrane Preparation is an ideal tool for screening for compounds binding to ET_B. 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 ET_B Membrane Prep yields greater than 5 fold signal-to-background ratio.

References

Product Information
Format	Membranes
Presentation	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.

Product Information

Format

Membranes

Presentation

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.

Applications
Application	Human ETB GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications	Radioligand binding assay GTPγS binding
Application Notes	Radioligand binding assay and GTPγS binding

Biological Information
Entrez Gene Number	NM_00015
Gene Symbol	EDNRA ETA ETRA
GPCR Class	A
Host Cells	CHO (Chinese hamster ovary cells)
Protein Target	ETB
Target Sub-Family	Endothelin
UniProt Number	P24530
Incubation 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.

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

_B

Table 1. Signal:background and specific binding values obtained in a competition binding assay with ET membrane prep.

Product Usage Statements

	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 Usage Statement

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.

Storage and Shipping Information
Storage Conditions	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.