Ast alt assay kit

Liver tissues (all lobes) were fixed in 10% formalin overnight, and then dehydrated and embedded in paraffin blocks, which were sectioned at a thickness of 5 μm. For Sirius red staining, sections were stained with hematoxylin (Weigert’s) for 8 min, washed with running tap water for 10 min, followed by incubation with 0.1% (wt/vol) Sirius red diluted in picric acid solution for 1 h. The slides were then rinsed in two quick changes of 0.5% (vol/vol) acetic acid to remove unbound dye. For immunostaining, sections were first heated in sodium citrate buffer to retrieve antigen. Sections were then blocked in 10% FBS blocking buffer followed by incubation with the following primary antibodies: anti-c-Jun (60A8; Cell Signaling), anti-F4/80 (SP115; Thermo Fisher Scientific), and anti-Ki67 (15580; Abcam), before quenching endogenous peroxidase using 3% hydrogen peroxide. TUNEL staining was performed as per manufacturer’s recommendations using the apoptosis detection kit (206386; Abcam). To quantify staining, 20 randomly taken images of 10× fields per section were evaluated by Meta Imaging Series (Molecular Devices) software.
A small portion of the liver was homogenized in ice cold NaCl buffer. The supernatant was collected, normalized before subjected to AST/ALT measurements using an AST/ALT Assay kit (Nanjing Jiancheng Bioengineering Institute) according to the supplier’s protocol.