|Description:||The ERR (estrogen related receptor) alpha TR-FRET Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential ERR alpha ligands as inverse agonists (sometimes referred to as antagonists) of coactivator displacement. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and an ERR alpha ligand-binding domain (ERR alpha-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous mix-and-read assay format.
In the ERR alpha TR-FRET Coactivator Assay, ERR alpha-LBD is added to ligand test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the ERR 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 displace coactivator peptide.
|Applications:||Co-Factor Interaction Assay|
|Storage:||Store in ultra-cold freezer (-68 to -85°C).|
|Size:||800 x 20 µL assays|
|Kit Components:||The TR-FRET ERR alpha Coactivator Assay Kit contains ERR alpha-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).|
|Materials Required but Not Supplied:||Microplate Reader|