LanthaScreen™ TR-FRET ER alpha Coactivator Assay Kit (Thermo Scientific™)

The LanthaScreen™ TR-FRET ER alpha Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential estrogen receptor (ER) alpha ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses an ER alpha ligand-binding domain (ER alpha-LBD) 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 alpha Coactivator Assay in agonist mode (to identify agonist compounds), ER alpha-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-anti-GST antibody. After incubation at room temperature, the TR-FRET 520:495 nm emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based on the biology of the ER alpha-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
To run the LanthaScreen TR-FRET ER alpha Coactivator Assay in antagonist mode (to identify antagonist compounds), ER alpha-LBD is added to ligand test compounds, followed by addition of a mixture of agonist, fluorescein-coactivator peptide, and terbium (Tb)-labeled 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.

Order Codes

Code Description
A15885 Catalog Number: A15885
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