ChemiScreen™ Membrane Preparation Human Recombinant EP2

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ChemiScreen™ Membrane Preparation Human Recombinant EP2

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货号：HTS185M

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	Chem-1	EP2

Description
Catalogue Number	HTS185M
Trade Name	ChemiScreen Chemicon
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). 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.

References

Product Information
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 1 mg/mL in packaging buffer, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80°C.

Applications
Application	Human EP2 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_000956.2
Gene Symbol	PTGER2 EP2
GPCR Class	A
Host Cells	Chem-1
Protein Target	EP2
Target Sub-Family	Prostanoid
UniProt Number	P43116
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.

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

₂ Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of EP Receptor membrane prep.

Product Usage Statements

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

1 unit = 10 µg
Bmax for [ ³H]-PGE ₂ binding: 4.38 pmol/mg protein
Kd for [ ³H]-PGE ₂ 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
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.