Principle of Assay
This kit is a more direct detection method for detecting the opening of the mitochondrial permeability transition pore (MPTP) than that based only on mitochondrial membrane potential analysis. The principle is as follows. Firstly, loading Calcein AM through passive transportation to fluorescently label living cells. It can easily penetrate the living cell membrane and is cleaved by intracellular esterase to form the membrane-impermeable polar molecule Calcein, which is then trapped in the cell and causes the cytoplasm including mitochondria to emit strong green fluorescence. After adding cobalt chloride, the fluorescence from the cytoplasm is quenched by cobalt chloride, leaving only fluorescence in the mitochondria. As a control, cells can be loaded with Calcein AM and cobalt chloride, and treated with ionomycin, so as to make cells load more Ca2+, which causes the activation of MPTP and quenching of mitochondrial fluorescence.
