LanthaScreen® TR-FRET ER beta Coactivator Assay Kit

Catalog number: A15893

 Related applications: Target & Lead Identification & Validation

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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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 800 x 20 µL assays
Validated Application: Co-Factor Interaction Assay, Nuclear Receptor Assay

Contents & storage

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)

Documents

Manuals & protocols