CP-640186 hydrochloride (CP 640186 HCl) is an isozyme-nonselective acetyl-CoA carboxylase (ACC) inhibitor, including rat liver ACC1 (IC50: 53 nM) and rat skeletal muscle ACC2 (IC50: 61 nM); with higher metabolic stability than CP-610431.

ACC1:53 nM, ACC2:61 nM

Aconitase activities are determined by homogenization of mouse brain tissues on ice at 10% w/v in CellLytic MT Mammalian Tissue Lysis/Extraction buffer, followed by centrifugation at 800×g?for 10?min at 4°C. Tissue lysates (50?μL) are then added to 200?μL of substrate mix (50?mM Tris/HCl pH 7.4, 0.4?mM NADP, 5?mM Na citrate, 0.6?mM MgCl2, 0.1% (v/v) Triton X-100 and 1U isocitrate dehydrogenase) and the reactions are incubated at 37°C for 15?min, followed by spectrophotometric absorbance measurements every minute for 15?min at 340?nm 37°C to determine the reaction slope. Aconitase activities of mouse brain tissues are then normalized to citrate synthase activities, which are determined using a citrate synthase assay kit.