ChemiSCREEN Membrane Preparation Recombinant Human EP₂ Prostanoid Receptor

产品说明

一般描述

Prostanoids bind to a family of 8 GPCRs to exert their biological effects (Narumiya and FitzGerald, 2001). The protanoid PGE₂ causes pain, vasodilation, immunosuppression of T cells, bone resorption and promotion of carcinogenesis. Four related GPCRs, EP₁, EP₂, EP₃ and EP₄, each bind to PGE₂, but the different G protein coupling status of each receptor leads to distinct biological effects. EP₂ couples primarily to Gs to increase intracellular cAMP levels. Mice deficient in EP₂ receptor showed impaired ovulation and fertilization, salt-sensitive hypertension (Kennedy et al., 1999). It has been shown that EP₂ receptors are also involved in cancer associated immunodeficiency. Thus, genetic knockout of the EP₂ receptor reduced tumor growth and prolonged survival in mice that had undergone isograft injection of MC26 or Lewis lung carcinoma cells (Yang et al., 2003). Millipore′s EP₂ 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 agonists and antagonists of EP₂. The membrane preparations exhibit a Kd of 14.52 nM for [³H]-PGE₂. With 7.5 nM [³H]-PGE₂, 10µg/well EP₂ Membrane Prep typically yields greater than 3-fold signal-to-background ratio.
Full-length human PTGER2 cDNA encoding EP₂

应用

Human EP2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

生化/生理作用

Target Sub-Family: Prostanoid
Protein Target: EP2
GPCR Class: A

质量

Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of EP₂ Receptor membrane prep.

	10 µg/well
Signal:Background	3.60
Specific Binding (cpm)	1146

1 unit = 10 µg
Bmax for [³H]-PGE₂ binding: 4.38 pmol/mg protein
Kd for [³H]-PGE₂ binding: ~14.52 nM

产品规格

Inucbation 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, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 500mM NaCl. 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, filtered and stored at 4°C.
Radioligand: [³H]-PGE₂ (Perkin Elmer#: NET-428)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, filtered and stored at 4°C.

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.41

产品性质

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_000956.2
UniProt登记号	P43116
运输	dry ice
Gene Information	human ... PTGER2(5732)

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