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HTS142M ChemiScreen™ Membrane Preparation
Recombinant Human EP4 Prostanoid Receptor

Human EP4 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P35408 Entrez Gene Number: NM_000958.2
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HTS142M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1EP4
      Description
      Catalogue NumberHTS142M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Membrane Preparation
      Recombinant Human EP4 Prostanoid Receptor
      OverviewHuman EP4
      Background InformationProstanoids are a series of arachidonic acid metabolites produced by the action of cyclooxygenase and further modified by isomerases and syntheses. Cells rapidly secrete prostanoids after synthesis, whereupon the prostanoids bind to a family of 8 GPCRs to exert their biological effects (Narumiya and FitzGerald, 2001). The prostaglandin PGE2 causes pain, vasodilation, immunosuppression of T cells, bone remodeling 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. EP4 couples primarily to Gs to increase intracellular cAMP levels. During neonatal development, EP4 participates in closure of the ductus arteriosus, a process required for switching circulation from the placenta to the lungs (Nguyen et al., 1997). In addition, EP4 mediates PGE2-induced bone formation by promoting osteoblastogenesis, and selective EP4 agonists are being evaluated as potential treatments for osteoporosis (Yoshida et al., 2002). Chemicon's EP4 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 antagonists of EP4 interactions with prostaglandin E2. The membrane preparations exhibit a Kd of 1.27-1.85 nM for [3H]-Prostaglandin E2. With 5 µg/well EP4 Membrane Prep and 1.5 nM [3H]-Prostaglandin E2, a greater than 3-fold signal-to-background ratio was obtained.
      References
      Product Information
      FormatMembranes
      PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 3-fold signal:background with 3H-labeled prostaglandin E2 at 1.5 nM.

      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 buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman EP4 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay, and GTPγS binding.
      Biological Information
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a member of the G-protein coupled receptor family. This protein is one of four receptors identified for prostaglandin E2 (PGE2). This receptor can activate T-cell factor signaling. It has been shown to mediate PGE2 induced expression of early growth response 1 (EGR1), regulate the level and stability of cyclooxygenase-2 mRNA, and lead to the phosphorylation of glycogen synthase kinase-3. Knockout studies in mice suggest that this receptor may be involved in the neonatal adaptation of circulatory system, osteoporosis, as well as initiation of skin immune responses.
      Gene Symbol
      • PTGER4
      • MGC126583
      • EP4
      • EP4R
      • PTGER2
      GPCR ClassA
      Host CellsChem-1
      Protein TargetEP4
      Target Sub-FamilyProstanoid
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P35408 # Receptor for prostaglandin E2 (PGE2). The activity of this receptor is mediated by G(s) proteins that stimulate adenylate cyclase. Has a relaxing effect on smooth muscle. May play an important role in regulating renal hemodynamics, intestinal epithelial transport, adrenal aldosterone secretion, and uterine function.
      SIZE: 488 amino acids; 53119 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: High in intestine and in peripheral blood mononuclear cells; low in lung, kidney, thymus, uterus, vasculature and brain. Not found in liver, heart, retina oe skeletal muscle.
      SIMILARITY: SwissProt: P35408 ## Belongs to the G-protein coupled receptor 1 family.
      Incubation ConditionsMembranes 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, 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: [3H] prostaglandin E2 (Perkin Elmer # NET428)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      SPECIFICATIONS:
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of EP4 membrane prep:
      Product Usage Statements
      10 µg/well
      Signal:Background 3.9
      Specific Binding (cpm) 643


      1 unit = 10 µg membrane preparation
      Bmax: 16.1 pmol/mg
      Kd: 1.57 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 ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      HTS142M 04053252325236

      Documentation

      ChemiScreen™ Membrane Preparation
      Recombinant Human EP4 Prostanoid Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiScreen™ Membrane Preparation
      Recombinant Human EP4 Prostanoid Receptor Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN EP4 PROSTANOID RECEPTOR - 2188021 2188021

      References

      Reference overviewPub Med ID
      Stimulation of bone formation and prevention of bone loss by prostaglandin E EP4 receptor activation
      Yoshida, Keiji, et al
      Proc Natl Acad Sci USA, 99:4580-5 (2002)  2002

      		11917107 11917107
      Genetic and pharmacological analysis of prostanoid receptor function
      Narumiya, S and FitzGerald, G A
      J Clin Invest, 108:25-30 (2001)  2001

      		11435452 11435452
      The prostaglandin receptor EP4 triggers remodelling of the cardiovascular system at birth
      Nguyen, M, et al
      Nature, 390:78-81 (1997)  1997

      		9363893 9363893

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