| Solution Information | help | |
| Enzyme: | Peroxisome proliferator-activated receptor gamma | |
| inhibitor: | BDBM79443 | |
| 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 dose response curves for compounds identified as possible PPAR gamma partial agonist probe candidates. In this assay, Cos-1 cells co-transfected with a full length PPARgamma construct in a pSport6 vector backbone (pS6-hPPARg) and three copies of a PPARg response element (3x-PPRE)-Aromatase-luciferase reporter construct, are incubated for 20 hours with test compound. As designed, a compound that activates PPARg activity will bind and activate the pS6-PPARg construct, thereby stimulating PPARg-mediated activation of the 3xPPRE-Aromatase-luciferase reporter, leading to an increase in well luminescence. Compounds are tested in triplicate using an 8-point titration series starting at a nominal concentration of 5 uM (range 5 uM to 0.002 uM). Protocol Summary: Luciferase reporter assays were conducted using a pSport-hPPARg full length construct and 3x PPRE luciferase reporter cotransfected into Cos-1 cells. Reverse transfections we | |
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