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HTS099M ChemiSCREEN™ Membrane Preparation Recombinant Human EP1 Prostanoid Receptor

Human EP1 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: P34995 Entrez Gene Number: NM_000955
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HTS099M
200 units  
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      Overview

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1EP1
      Description
      Catalogue NumberHTS099M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human EP1 Prostanoid Receptor
      OverviewFull-length human PTGER1 cDNA encoding EP1
      Background InformationProstanoids are a series of arachidonic acid metabolites produced by the action of cyclooxygenase and subsequently by isomerases and synthases. 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 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; EP1 couples primarily to Gq to mobilize intracellular calcium. EP1 appears to mediate the effects of PGE2 in promoting formation of precancerous lesions in animal models of colon cancer (Watanabe et al., 1999). In addition, EP1 has an inhibitory effect on stress-induced aggressive and risk-taking behaviors in mice (Matsuoka et al., 2005). Millipore's EP1 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 EP1. The membrane preparations exhibit a Kd of 14.5 nM for [3H]-PGE2. With 40 nM [3H]-PGE2, 10 μg/well EP1 Membrane Prep typically yields greater than 3-fold signal-to-background ratio.
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
      Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman EP1 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 SummaryThis gene encodes one of two high affinity forms of a receptor for melatonin, the primary hormone secreted by the pineal gland. This gene product is an integral membrane protein that is a G-protein coupled, 7-transmembrane receptor. It is found primarily in the retina and brain although this detection requires RT-PCR. It is thought to participate in light-dependent functions in the retina and may be involved in the neurobiological effects of melatonin.
      Gene Symbol
      • PTGER1
      GPCR ClassA
      Host CellsChem-1
      Protein TargetEP1
      Target Sub-FamilyProstanoid
      UniProt Number
      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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 10mM MES, pH 6, 1mM EDTA, 10mM MnCl2. 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: 10mM MES, pH 6, 1mM EDTA, 10mM MnCl2, filtered and stored at 4°C

      Radioligand: [3H]- PGE2. (Perkin Elmer # NET-428 )

      Wash Buffer: 10mM MES, pH 6, 1mM EDTA, 10mM MnCl2, filtered and stored at 4°C.

      One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 3-fold signal: background with 3H labeled PGE2.
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with EP1 membrane prep.
      Product Usage Statements
      10 µg/well
      Signal:Background 4.3
      Specific Binding (cpm) 724

      SPECIFICATIONS: 1 unit = 10 μg membrane preparation
      Bmax: 4.6 pmol/mg
      Kd: 14.5 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 ConditionsStore 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
      Material Size200 units
      Global Trade Item Number
      Catalogue Number GTIN
      HTS099M 04053252588686