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HTS176M ChemiSCREEN™ Membrane Preparation Recombinant Human S1P1 Lysophospholipid Receptor

Human S1P1 / EDG1 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: P21453 Entrez Gene Number: NM_001400
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HTS176M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1S1P1 / EDG1
      Description
      Catalogue NumberHTS176M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human S1P1 Lysophospholipid Receptor
      OverviewFull-length human EDG1 cDNA encoding S1P1
      Background InformationSphingosine 1-phosphate (S1P) is a biologically active lysophospholipid that transmits signals through a family of five G-protein-coupled receptors to regulate cell proliferation, migration, cytoskeletal organization, and differentiation. S1P1 was the first identified S1P receptor. It primarily couples to PTX-sensitive Gi/o proteins and mediates S1P-induced adenylate cyclase inhibition. Expression of S1P1 is pervasive, including spleen, brain, heart, lung, adipose tissues, liver, thymus, kidney, and skeletal muscle (Zhang et al. 1999). The deletion of S1P1 in mice results in embryonic lethality (Liu et al., 2000) with death attributable to incomplete vascular maturation. Recent reports demonstrate specific roles for S1P1 in lymphocyte recirculation/egress (Matloubian et al., 2004). Millipore's S1P1 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 S1P1 interaction with its ligands. The cell line exhibits a calcium response with EC50s of 141 μM for Sphingosine 1-phosphate. The membrane preparations exhibit EC50s of 9.5 μM for Sphingosine 1-phosphate in a GTPγS binding assay.
      References
      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 1 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman S1P1 / EDG1 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
      Gene Symbol
      • S1PR1
      • CHEDG1
      • EDG1
      GPCR ClassA
      Host CellsChem-1
      Protein TargetS1P1 / EDG1
      Target Sub-FamilyLysophospholipid
      UniProt Number
      Incubation ConditionsMembranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [35S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl2/10 μM GDP in a nonbinding 96-well plate. Unlabeled Sphingosine 1-phosphate was added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (Millipore MAHF B1H) previously prewetted with water. The plate is washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4, then dried and counted.
      Note: Performing the reaction with lower GDP concentrations (0.5 μM) results in elevated constitutive activity (Figure 2).

      One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific sodium propionate-stimulated [35S]-GTPγS binding.

      The S1P1 membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assurance1 unit = 5 μg
      EC50 in GTPγS binding assay by Sphingosine 1-phosphate: ~ 9.5 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 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
      Catalog Number GTIN
      HTS176M 04053252581120

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      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations