|Description:||The TR-FRET FXR (farnesoid X receptor) Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential FXR ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and an FXR ligand-binding domain (FXR-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous, mix-and-read assay format.
When running the TR-FRET FXR coactivator assay in agonist mode, FXR-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 used to determine the EC50 from a dose response curve of the compound. Based on the biology of the FXR-coactivator peptide interaction, the ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide.
|Applications:||Nuclear Receptor Assay|
|Storage:||Tb-anti-GST Antibody (Rabbit): Store at -20 °C
FXR-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
|Size:||800 x 20 µL assays|
|Materials Required but Not Supplied:||Microplate Reader|