Solution Informationhelp
Enzyme: Peroxisome proliferator-activated receptor gamma
inhibitor: BDBM79439
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: Assay Overview: The purpose of this assay is to confirm compounds that can directly bind to PPARg through competition with a fluorescently labeled high affinity PPARg compound. The fluorescent ligand when bound to the PPARg LBD protein is in close proximity to the Tb-anti PPARg antibody bound to the N-terminal His tag on the PPARg LBD. In the absence of test compound, this provides a robust TR-FRET signal which is the ratio of the fluorescein emission at 520 nm and the Tb emission at 490 nm. When test compound displaces the fluorescently labeled control compound, it causes a loss of the TR-FRET signal which is proportional to how much of the compound is displaced. This assay allows for the separation of compounds positive in the cell-based luminescence assays that are working through direct binding to PPARg versus compounds modulating PPARg transactivation activity through indirect mechanisms. In addition, it provides a more sensitive measurement of compound binding to PPARg than the
 
 
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