Summary: The PPARγ (Peroxisome Proliferator-Activated Receptor-gamma) Competitor Assay, Green uses the human-derived recombinant PPARγ ligand-binding domain (PPARγ-LBD) tagged with an N-terminal GST-tag and a novel, tight-binding, selective fluorescent PPARγ ligand (PPARγ Green) in a homogenous, mix-and-read assay format. PPARγ-LBD/PPARγ Green complex produces high polarization. This complex is then added to individual test compounds in microwell plates. Competitors displace the fluorescent PPARγ Green Ligand from the PPARγ-LBD/PPARγ Green complex, causing the fluorescent ligand to tumble rapidly during its fluorescence lifetime, resulting in a low polarization value. Noncompetitors will not displace the fluorescent ligand from the complex, so the polarization value remains high. The shift in polarization value in the presence of the test compounds is used to determine relative affinity of test compounds for the PPARγ-LBD.
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