Fam flica in vitro caspase detection kit

According to the manufacturer’s instructions, caspase activity was measured using the FAM-FLICA in vitro caspase detection kit (ImmunoChemistry Technologies, Bloomington, MN, USA). Cells were harvested 24 h after PA and skatole treatment and cultured in a 5% CO₂ at 37 °C. After treatment, the pellet was re-suspended in apoptosis wash buffer, and the cell suspension (1 × 10⁵ cells) was transferred to a new tube. In the dark, the cell suspension was incubated with 1x FAM-FLICA caspase 3&7 for 1 h at 37 °C. Caspase activity was detected using an EnSpire multimode plate reader (PerkinElmer) at 488 nm emission/515 nm excitation.

To investigate apoptosis, caspases 3 and 7 were detected with the FAM-FLICA in vitro Caspase Detection Kit (ImmunoChemistry TECHNOLOGIES, Bloomington, MN, USA).

Apoptosis and necrosis assay were examined with a FAM-FLICA in vitro Caspase Detection Kit (ImmunoChemistry Technologies, Bloomington, IN, USA). Briefly, cultured cells were seeded onto 35-mm glass-coated wells (Iwaki, Chiba, Japan), incubated with FLICA at 39°C for 2 h, replated in fresh media, and then incubated again at 39°C for 1 h. The stained cells were incubated with Hoechst33258 and propidium iodide at room temperature for several minutes, fixed, and visualized with an epifluorescence microscope (Olympus, Tokyo, Japan).