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HTS092M | ChemiSCREEN™ EP3 Prostanoid Receptor Membrane Preparation

200 assays  
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      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1 EP3
      Catalogue NumberHTS092M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ EP3 Prostanoid Receptor Membrane Preparation
      OverviewFull-length human PTGER3 cDNA encoding splice variant 6 of EP3
      Background InformationProstanoids 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 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. Further diversity is generated by alternative splicing; the human gene for EP3 generates 9 alternatively spliced mRNAs encoding 8 isoforms of EP3 (Kotani et al., 1997). These isoforms of EP3 vary in sequence at their C-termini, and differ in their ability to couple to Gs, Gq or Gi (Kotani et al., 1995). EP3 is required for fever induced by pyrogens, a response long attributed to prostaglandins by the antipyretic action of aspirin and other COX inhibitors (Ushikubi et al., 1998). Chemicon's EP3 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 EP3 interactions and its ligands. The membrane preparations exhibit a Kd of 1.56 nM for [3H]-PGE2. With 1.5 nM [3H]-PGE2, 5 µg/well and 10 µg/well EP3 Membrane Prep typically yield greater than 10 and 20-fold signal-to-background ratios, respectively.
      Product Information
      PresentationOne package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 3H-labeled PGE2 at 1.5 nM

      Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
      Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
      ApplicationHuman EP3 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay and GTPS 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 may have many biological functions, which involve digestion, nervous system, kidney reabsorption, and uterine contraction activities. Studies of the mouse counterpart suggest that this receptor may also mediate adrenocorticotropic hormone response as well as fever generation in response to exogenous and endogenous stimuli. Multiple alternatively spliced transcript variants encoding eight distinct isoforms have been reported.
      Gene Symbol
      • PTGER3
      • EP3-III
      • EP3-I
      • PGE2-R
      • MGC141829
      • EP3-II
      • EP3
      • EP3e
      • MGC27302
      • MGC141828
      • EP3-IV
      GPCR ClassA
      Host CellsChem-1
      Protein TargetEP3
      Target Sub-FamilyProstanoid
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P43115 # Receptor for prostaglandin E2 (PGE2); the EP3 receptor may be involved in inhibition of gastric acid secretion, modulation of neurotransmitter release in central and peripheral neurons, inhibition of sodium and water reabsorption in kidney tubulus and contraction in uterine smooth muscle. The activity of this receptor can couple to both the inhibition of adenylate cyclase mediated by G-I proteins, and to an elevation of intracellular calcium. The various isoforms have identical ligand binding properties but can interact with different second messenger systems (By similarity).
      SIZE: 390 amino acids; 43310 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed at least in small intestine, heart and pancreas.
      SIMILARITY: SwissProt: P43115 ## 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
      Radioligand: [3H] PGE2 (Perkin Elmer#:NET-428 )
      Wash Buffer: 50 mM Tris, pH 7.4 filtered and stored at 4°C.
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of EP3 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 38 21
      Specific Binding (cpm) 6,386 4,053

      1 unit = 10 μg
      Bmax: 15.0 pmol/mg
      Kd: 1.56 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 assays
      Transport Information
      Supplemental Information




      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number


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

      11435452 11435452
      Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP3
      Ushikubi, F, et al
      Nature, 395:281-4 (1998) 1998

      9751056 9751056
      Structural organization of the human prostaglandin EP3 receptor subtype gene (PTGER3)
      Kotani, M, et al
      Genomics, 40:425-34 (1997) 1997

      9073510 9073510
      Molecular cloning and expression of multiple isoforms of human prostaglandin E receptor EP3 subtype generated by alternative messenger RNA splicing: multiple second messenger systems and tissue-specific distributions
      Kotani, M, et al
      Mol Pharmacol, 48:869-79 (1995) 1995

      7476918 7476918

      Data Sheet


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