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HTS068M ChemiSCREEN™ Membrane Preparation Recombinant Human APJ Apelin Receptor

Human APJ 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: P35414 Entrez Gene Number: NP_005152
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HTS068M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1APJ
      Description
      Catalogue NumberHTS068M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human APJ Apelin Receptor
      OverviewFull-length human AGTRL1 cDNA encoding APJ
      Background InformationApelin peptides have been discovered to be a family of peptides of different sizes that is derived from the N-terminus of a 77 amino acid precursor peptide (preproapelin) (Hosoya et al., 2000). Apelin receptor (APJ) is a G protein-coupled receptor that is activated by several apelin fragments. APJ activation leads to the inhibition of cAMP production. APJ and apelin peptides have been found to be involved in the regulation of cardiovascular function (Katugampola et al., 2001) and fluid homeostasis (Reaux et al., 2001). Broad roles of apelin system has been established in lowering blood pressure, as a potent cardiac inotrope, in modulating pituitary hormone release and food and water intake, in stress activation, and as a novel adipokine that is excreted from fat cells and regulates insulin (Lee et al., 2006) . Millipore's APJ 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 APJ. The membrane preparations exhibit a Kd of 0.4 nM for [125I]-Apelin. With 0.25 nM [125I]-Apelin, 5 g/well APJ Membrane Prep typically yields greater than 20-fold signal-to-background ratio.
      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 the indicated concentration in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman APJ 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
      • APLNR
      • AGTRL1
      • APJ
      GPCR ClassA
      Host CellsChem-1
      Protein TargetAPJ
      Target Sub-FamilyApelin
      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 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: [125I]-Apelin. (Perkin Elmer#:NEX-393 )

      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, 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 20-fold signal:background with 125I labeled Apelin at 0.25 nM
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of APJ membrane prep.
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 33 50
      Specific Binding (cpm) 17297 15015


      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax for [125I]-Apelin binding: 3.85 pmol/mg protein
      Kd for [125I]-Apelin binding: ~0.4 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
      HTS068M 04053252284762

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      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations