This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15135:
The LanthaScreen™ TR-FRET TR (thyroid receptor) beta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential TR beta ligands as agonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant TR beta ligand-binding domain (TR beta-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous, mix-and-read assay format.
To Assay: When performing the LanthaScreen™ TR-FRET TR beta coactivator assay, TR beta-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb 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 TR beta-coactivator peptide interaction, the ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, induce a conformational change, and recruit coactivator peptide (Figure 1).
Contents and Storage: The LanthaScreen™ TR-FRET TR beta Coactivator Assay Kit contains TR beta-LBD (GST) protein, fluorescently labeled SRC2-2 coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).
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