Millipore Sigma Vibrant Logo

CYL4039 PrecisION™ hERG ion channel membrane preparation

Human ERG ion channel membrane preparation for Radioligand binding Assays.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: 293 cell line Entrez Gene Number: U04270
View Products on Sigmaaldrich.com
CYL4039
200 units  
无法检索价格。
最小订购数量为倍数
Maximum Quantity is
以订单确认为准 更多信息
您节约了 ()。
 
—
Request Pricing
—
目前缺货 目前缺货
有货 
停产
少量现货
供货情况有待确认
    其余：另行通知
      其余：另行通知
      另行通知
      联系客户服务
      Contact Customer Service

       

      联系客户服务

      概述

      重要规格表

      SpeciesHost CellsProtein Target
      Human 293 cell linehERG
      Description
      Catalogue NumberCYL4039
      Trade Name
      • PrecisION
      DescriptionPrecisION™ hERG ion channel membrane preparation
      OverviewHuman ERG
      Background InformationThe human ether-a-go-go related gene (hERG) is a potassium ion channel which is essential for normal cardiac repolarization. In drug screening models, the hERG K+ channel has been indicated to inhibit a wide variety of compounds, and its blockage can lead to cardiac QT interval prolongation and life threatening arrhythmias (Murphy et al. 2006). Cardiac safety relating to IKr K+ channels has become a major concern of regulatory agencies, as hERG channel inhibition has been identified as the firmest link to QT prolongation (Chiu et al. 2004). Millipore's hERG membrane preparations are crude membrane preparations made from HEK293 stable recombinant cell lines (Millipore cat. # CYL3039), which are ideal HTS tools for screening antagonists against the hERG channel. The membrane preparations exhibit a Kd of 5.2 nM for [3H]-Astemizole. With 10 μg/well hERG Membrane Prep and 3.0 nM [3H]-Astemizole, a greater than 4-fold signal-to-background ratio was obtained.
      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 ERG ion channel membrane preparation for Radioligand binding Assays.
      Application NotesRadioligand binding assay
      Biological Information
      Entrez Gene Number
      Host Cells293 cell line
      Protein TargethERG
      Target Sub-FamilyPotassium
      Incubation ConditionsRECOMMENDED ASSAY CONDITIONS: Membranes 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 FB 96-well harvest plate (Millipore cat. # MAHF B1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 25 mM Tris, pH 7.4, 130 mM NaCl, 5 mM KCl, 0.8 mM CaCl2, 0.1% BSA. Binding reaction is transferred to the filter plate, and washed 6 times (250μL per well per wash) with Wash Buffer. The plate is dried and counted.

      Binding buffer: 10 mM Hepes, pH 7.4, 130 mM NaCl, 5 mM KCl, 0.8 mM MgCl2, 1 mM NaEGTA, 10 mM glucose, 0.1% BSA, filtered and stored at 4°C

      Radioligand: [3H]-Astemizole (Perkin Elmer # NET-1140)

      Wash Buffer: 25 mM Tris, pH 7.4, 130 mM NaCl, 5 mM KCl, 0.8 mM MgCl2, 0.05 mM CaCl2, 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 4-fold signal: background with 3H-labeled Astemizole.
      Cell Line Type
      • GPCR Cell Lines
      Table 1. Signal: background and specific binding values obtained in a competition binding assay for the hERG membrane prep.
      Product Usage Statements
      10 µg/well
      Signal:Background 4.6
      Specific Binding (cpm) 435


      SPECIFICATIONS: 1 unit = 10 μg membrane preparation with [3H]-astemizole
      Bmax with [3H]-astemizole: 6.7 pmol/mg
      Kd for [3H]-astemizole: 5.2 nM

      Table 2. Rank order comparison of various small molecule hERG inhibitors using [3H]-astemizole binding assay and E-Phys assays:
      Binding Ki (nM) with hERG membrane preps Electrophysiology IC50 (nM) with hERG cell lines
      Millipore CYL4039M Literature value* Millipore CYL4039 by IonWorks PPC Millipore CYL4039 by PatchExpressLiterature value by Patch Clamp
      Astemizole 3.4 ± 0.7 3.3 ± 0.7 4.6 ± 0.003 ND 0.5
      Dofetilide 5.8 ± 2.428 ± 6NDND15.3 ± 2.5
      E-4031 6.1 ± 0.8 58 ± 5115 ± 0.02 144 ± 44 14
      Pimozide 18.8 ± 2.3 14 ± 4NDND18
      Cisapride 104.9 ± 30.9 123 ± 25 75.6 ± 0.02 5 ± 1 44.5 ± 10.6
      Haloperidol 241.4 ± 21.4 234 ± 20 NDND63
      Risperidone > 4000 4302 ± 319NDND167
      Verapimil > 4000 3902 ± 529 1332 ± 1203100 ± 400 830

      *Data obtained from Chiu et al., 2004


      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 up to expiration date indicated on the label. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Global Trade Item Number
      产品目录编号 GTIN
      CYL4039 04053252672682