GW1929 Inhibitor

Solid. Powder

98.00%

Store at -20 degree C for 3 years powder
-80 degree C for 2 years in solvent

495.57

DMSO: 33mg/mL (66.59mM)
(<1mg/ml refers to the product slightly soluble or insoluble)

C30H29N3O4

CN(CCOc1ccc(C[C@H](Nc2ccccc2C(=O)c2ccccc2)C(O)=O)cc1)c1ccccn1

196808-24-9

For the experiments, the cells are plated in 96-well plates at a density of 2 x 10^5 cells per cm2 and cultured in the presence of TBBPA, in concentrations ranging from 1nM to 100uM TBBPA. TBBPA is dissolved in DMSO, resulting in a final vehicle concentration of 0.1% (v/v). Control (no vehicle) and DMSO-treated wells are included in the experimental design to determine the effect of DMSO. To study whether PPAR-gamma is involved in the neurotoxic effect of TBBPA, cells are co-treated with 10uM TBBPA and 10uM GW1929 or GW9662. After 6 or 24 hours of culture, 100uL medium is collected for the LDH analysis, and the cells are collected and frozen at -70 degree C for the caspase-3 activity measurements [2].

Ligand binding to bacterially expressed ligand-binding domain (LBD) of hPPAR-gamma is determined by the scintillation proximity assay (SPA). The assay measures the ability of putative ligands to displace receptor-bound [3H]BRL 49653. Assays are conducted in 96-well plates. Wells contained varying concentrations of GW1929 or troglitazone; streptavidin-modified SPA beads to which biotinylated PPAR-gamma LBD is prebound; and 10nM of the specific radioligand [3H]BRL 49653 in a volume of 100 uL. The amount of nonspecific binding, as assessed by control wells that contained 50uM of the corresponding unlabeled ligand, is subtracted from each data point. For each compound tested, plots of ligand concentration versus counts/min of radioligand bound are constructed, and apparent Ki values are estimated from a nonlinear least-squares fit of the data, assuming simple competitive binding. The results are expressed as pKi, where pKi = -log10(KI) [1].

Metabolism; PPAR