Assay Method Information

Assay Name:  Competition Binding Assay (Ki)
Description:  The scintillation proximity assay was performed in 96-well plates containing polylysine-coated yttrium silicate beads, His-PPARgamma-LBD, and [3H]rosiglitazone. Test compounds were tested in 10-point concentration-response curves starting at the indicated concentration. All components were added simultaneously and incubated with gentle shaking for 1 h at room temperature. Scintillation counts were determined in a Microbeta 1450 Wallac Trilux counter (PerkinElmer Life Sciences), reading each well for 1 min. Wells devoid of competitor represented 100% binding. Nonspecific binding was measured by leaving PPARgamma protein out of the scintillation proximity assay reaction. Experiments were repeated three times. Ki values of the ligands were calculated using Graph-Pad Prism version 4.0c for Macintosh (GraphPad Software).
Affinity data for this assay

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