Key Specifications Table
Species | Host Cells | Protein Target |
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Human | Chem-1 | EP2 |
Description | |
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Catalogue Number | HTS185M |
Trade Name |
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Description | ChemiSCREEN™ Membrane Preparation Recombinant Human EP2 Prostanoid Receptor |
Overview | Full-length human PTGER2 cDNA encoding EP2 |
Background Information | 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). Millipores 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. |
Product Information | |
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Format | Membranes |
Presentation | Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives. Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffe, rapidly frozen, and stored at -80°C. |
Quality Level | MQ100 |
Applications | |
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Application | Human EP2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding. |
Key Applications |
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Application Notes | Radioligand binding assay and GTPγS binding |
Biological Information | |
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Entrez Gene Number |
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Gene Symbol |
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GPCR Class | A |
Host Cells | Chem-1 |
Protein Target | EP2 |
Target Sub-Family | Prostanoid |
UniProt Number |
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UniProt Summary | FUNCTION: SwissProt: P43116 # Receptor for prostaglandin E2 (PGE2). The activity of this receptor is mediated by G(s) proteins that stimulate adenylate cyclase. The subsequent raise in intracellular cAMP is responsible for the relaxing effect of this receptor on smooth muscle. SIZE: 358 amino acids; 39761 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. TISSUE SPECIFICITY: Placenta and lung. SIMILARITY: SwissProt: P43116 ## Belongs to the G-protein coupled receptor 1 family. |
Incubation 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. |
Product Usage Statements |
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10 µg/well | |
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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 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 | |
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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. |
Packaging Information | |
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Material Size | 200 units |