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 EC50from a dose response curve of the compound. Based on the biology of the ERR alpha-coactivator peptide interaction, this ligand EC50is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and displace coactivator peptide.
Storage :
Tb-anti-GST Antibody (Rabbit): Store at -20°C
ERR alpha-LBD, GST: Store at -80°C
Fluoroscein Coactivator Peptide: Store at -20°C
TR-FRET Coregulatore Buffer G: Store at 4°C
1M DTT: Store at -20 °C or -80°C
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