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LanthaScreen™ TR-FRET ER beta Coactivator Assay Kit

LanthaScreen™ TR-FRET ER beta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential estrogen receptor (ER) beta ligands as agonists or antagonists of ligand-dependent coactivator recruitment.

Manufacturer:  LanthaScreen™ A15893

Catalog No. A15893


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Description

Description

  • The LanthaScreen™ TR-FRET ER beta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential estrogen receptor (ER) beta ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses a ER beta ligand-binding domain (ER beta-LBD) that is tagged with glutathione-S-transferase (GST), a terbium (Tb)-labeled anti-GST antibody, and a fluorescein-labeled coactivator peptide in a homogenous mix-and-read assay format.
  • Agonist Mode - To run the LanthaScreen TR-FRET ER beta Coactivator Assay in agonist mode (to identify agonist compounds), ER beta-LBD is added to ligand test compounds followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-anti-GST antibody. After room temperature incubation, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based upon the biology of the ER beta-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide.
  • Antagonist Mode - When the LanthaScreen TR-FRET ER beta Coactivator Assay Kit is run in antagonist mode (to identify antagonist compounds), ER beta-LBD is added to ligand test compounds followed by the addition of a mixture of agonist, fluorescein-coactivator peptide, and Tb-anti-GST antibody. The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.
  • Includes:
    • 1 tube ER beta Ligand Binding Domain Recombinant Protein (store at -68 to -85°C)
    • 1 tube of Peptide (store at -5 to -30°C)
    • 2 bottles TR-FRET Buffer (store at -5 to -30°C)
    • 1 vial Tb-Anti-GST Ab (store at -5 to -30°C)
    • 1 tube of DTT (store at -5 to -30°C)
Specifications

Specifications

Co-Factor Interaction Assay, Nuclear Receptor Assay
800 x 20 μL assays
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