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| Solution Information | help | |
| Enzyme: | Isoform 2 of Nuclear receptor corepressor 2 (TRAC-1) | |
| inhibitor: | BDBM75979 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this biochemical assay is to determine whether compounds presenting an inverse agonist profile as active in a previous set of experiments entitled, TR-FRET-based primary biochemical high throughput screening assay to identify agonists of nuclear receptor subfamily 2, group E, member 3 (NR2E3) (AID 2300), were nonselective due to activation of the interaction between PPARgamma-NCOR2. This assay determines dose response curves for tested compounds. In this HTRF-format assay GST-PPARg and its interaction partner, a biotinylated corepressor NCOR2 are incubated in the presence of test compounds, Eu(K)-anti GST antibody, and Streptavidin-D2. Interaction between the NCOR2 and PPARg partners brings the fluorophores together, leading to well FRET. As designed, test compounds that act as PPARg inverse agonists will lead to strengthening of PPARg and corepressor NCOR2 interaction, thereby enhancing interaction of the fluorescent tags, leading to increased well FR | |
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