HTS185M
ChemiSCREEN Membrane Preparation Recombinant Human EP2 Prostanoid Receptor
Human EP2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
Code UNSPSC :
41106514
eCl@ss :
32161000
Nomenclature NACRES :
NA.41
Source biologique
human
Niveau de qualité
Produit recombinant
expressed in Chem-1 cells
Fabricant/nom de marque
ChemiScreen
Chemicon®
Technique(s)
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
Numéro d'accès NCBI
Numéro d'accès UniProt
Conditions d'expédition
dry ice
Informations sur le gène
human ... PTGER2(5732)
Description générale
Full-length human PTGER2 cDNA encoding EP2
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.
Application
Human EP2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Qualité
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of EP2 Receptor membrane prep.
1 unit = 10 µg
Bmax for [3H]-PGE2 binding: 4.38 pmol/mg protein
Kd for [3H]-PGE2 binding: ~14.52 nM
| 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
Caractéristiques
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.
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.
Informations légales
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Code de la classe de stockage
12 - Non Combustible Liquids
Classe de danger pour l'eau (WGK)
WGK 2
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
C R Kennedy et al.
Nature medicine, 5(2), 217-220 (1999-02-04)
Prostaglandins (PGs) are ubiquitous lipid mediators derived from cyclooxygenase metabolism of arachidonic acid that exert a broad range of physiologic activities, including modulation of inflammation, ovulation and arterial blood pressure. PGE2, a chief cyclooxygenase product, modulates blood pressure and fertility
Cancer-associated immunodeficiency and dendritic cell abnormalities mediated by the prostaglandin EP2 receptor
Yang, Li, et al
The Journal of Clinical Investigation, 111, 727-735 (2003)
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