GW6471

2 years -80 in solvent

0.24 uM [1], 0.24 uM [1], 0.24 uM [1], 0.24 uM [1], 0.24 uM [1], 0.24 uM [1]

Binding assays, The effects of GW6471 on the interaction of coactivator and co-repressor peptides with PPAR are determined by chemical-mediated fluorescence energy transfer assays. The experiments are conducted with 5 nM PPARalpha LBD of biotinylated peptide containing individual motifs , following the manufacturer's instructions for the hexahistidine detection kit in a buffer containing 50 mM MOPS, pH 7.4, 50 mM NaF, 0.05 mM CHAPS, 0.1 mg/mL bovine serum albumin, and 10 mM dithiothreitol (DTT). The binding signals are detected with the increasing concentrations of GW6471, and the results from four repeated experiments are normalized as a percentage of the binding in the absence of GW6471. The effects of GW6471 on the affinity of the SMRT or N-CoR peptides with purified PPAR LBD are determined by fluorescence polarization in a buffer containing 10 mM HEPES, pH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.005% polysorbate-20, 5 mM DTT and 2.5% DMSO. Varied concentration of PPAR LBD in the presence or absence of 40 uM GW6471 are incubated at room temperature with 10 nM of a fluorescein-labelled peptide of N-CoR2 or SMRT2. The fluorescence polarization values for each concentration of receptor are determined using a BMG PolarStar Galaxy fluorescence reader with 485 nm excitation and 520 nm emission filters. The apparent dissociation constant (Kd) values are determined by the binding curves derived from a nonlinear least-squares-fit of the data for a simple 1:1 interaction.

(S, Z)-N-(3-(4-(2-(5-methyl-2-phenyloxazol-4-yl)ethoxy)phenyl)-2-(4-oxo-4-(4-(trifluoromethyl)phenyl)but-2-en-2-ylamino)propyl)propionamide