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HTS063M ChemiSCREEN™ Membrane Preparation Recombinant Human GPR43 Free Fatty Acid Receptor

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HTS063M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1FFA2 / GPR43
      Description
      Catalogue NumberHTS063M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human GPR43 Free Fatty Acid Receptor
      OverviewFull-length human GPR43 cDNA
      Background InformationGPR43 is a GPCR that, along with GPR41, is activated by short chain carboxylic acids formate, acetate, proprionate, butyrate and pentanoate. Neutrophils and other leukocytes selectively express GPR43, and GPR43 mediates calcium flux and chemotaxis in these cells by coupling to both Gq and Gi (Brown et al., 2003; Le Poul et al., 2003). Millipore's GPR43 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 GPR43 interactions with its ligands. The cell line exhibits a calcium response with EC50s of 11.8μM for sodium propionate. The membrane preparations exhibit EC50s of 40.7 μM for sodium propionate 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: 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.
      Quality LevelMQ100
      Applications
      ApplicationHuman FFA2 / GPR43 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
      • FFAR2
      • FFA2
      • GPCR43
      • GPR43
      GPCR ClassA
      Host CellsChem-1
      Protein TargetFFA2 / GPR43
      Target Sub-FamilyFree Fatty Acid
      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 MgCl2/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled sodium propionate 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.

      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 GPR43 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 sodium propionate: ~ 40.7 μM
      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
      HTS063M 04053252732683

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      Categories

      Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations