| Solution Information | help | |
| Enzyme: | Peroxisome proliferator-activated receptor alpha | |
| inhibitor: | BDBM79449 | |
| 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 determine the selectivity of the probe ML215 for PPARG (gamma) over PPARA (alpha). In this assay, Cos-1 cells co-transfected with a Gal4 PPARA LBD construct in a pBind vector backbone (pBind-Gal4-PPARA-LBD) and a UAS response element (UAS)-luciferase reporter construct, are incubated for 20 hours with test compound. As designed, a compound that activates PPARA activity will bind and activate the pBind-Gal4-PPARA-LBD construct, thereby stimulating PPARA-mediated activation of the UAS- luciferase reporter, leading to an increase in well luminescence. Compounds were tested in triplicate using an 8-point titration series starting at a nominal concentration of 3 uM (range 3 uM to 0.001 uM). The probe, ML215, was not active against PPARA in this assay demonstrating its selectivity as a probe for PPARG. Protocol Summary: Luciferase reporter assays were conducted using a pBind-Gal4PPARaLBD construct and UAS luciferase reporter cotransfected i | |
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