|Description:||The TR-FRET PPAR (peroxisome proliferator activated receptor) delta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential PPAR delta 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 PPAR delta ligand-binding domain (PPAR delta-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous mix-and-read assay format.
When running the TR-FRET PPAR delta Coactivator Assay in agonist mode (to identify agonist compounds), PPAR delta-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-labeled coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the TR-FRET ratio of 520/495 is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PPAR delta-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.
|Applications:||Nuclear Receptor Assay|
|Storage:||Tb-anti-GST Antibody (Rabbit): Store at -20 °C
PPAR Delta-LBD, GST: Store at -80 °C
Fluoroscein Coactivator Peptide: Store at -20 °C
TR-FRET Coregulatore Buffer J: Store at 4 °C
1M DTT: Store at -20 °C or -80 °C
|Size:||800 x 20 µL assays|
|Kit Components:||The TR-FRET PPAR delta Coactivator Assay Kit contains PPAR delta-LBD (GST) protein, fluorescently labeled C33 coactivator peptide, Tb-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|