Mitochondrial Permeability Transition Pore (mPTP) Fluorometric Assay Kit

Fluorescence microscope;Flow cytometry

Fluorometric method

Cell-based (quantitative)

Mitochondria are the energy centers of cells and play a very important role in the processes of apoptosis and necrotic cell death. Mitochondrial permeability transition pore (mPTP), also known as mitochondrial megachannel (MMC), It is a non-specific channel composed of the inner and outer membranes of mitochondria, which participates in the release of substances within mitochondria during cell death. The opening of mitochondrial permeability transition pores is a key event causing cell death and plays an important role in the regulation of cell survival and apoptosis.
The detection principle of this kit: The kit uses Calcein AM to stain cells, causing the cytoplasm, including mitochondria, to show strong green fluorescence. Under physiological pH conditions, when Calcein complexes with Co²⁺, the fluorescence signal is quenched. Under normal circumstances, the mPTP in mitochondria is off, and at this time,Co²⁺ cannot enter the mitochondria, resulting in only green fluorescence appearing within the mitochondria. The inner mitochondrial membrane of normal cells can maintain a normal mitochondrial potential gradient to ensure cellular respiration and energy supply. With the intake and release of Ca²⁺, a low-conductivity permeability conversion pore switches back and forth between opening and closing. When cells undergo apoptosis and pathological death, the permeability of mitochondrial membrane potential conversion pores changes. Overload of Ca²⁺, oxidation of mitochondrial glutathione, increase in reactive oxygen species levels, including subsequent release of cytochrome C, and decrease in mitochondrial membrane potential can all lead to the activation of mitochondrial permeability conversion pores. Further treatment with the calcium ion carrier Ionomycin as a control to induce a large amount of extracellular Ca²⁺ to enter the intracellular and mitochondrial matrix and cause the opening of mPTP will weaken or disappear the green fluorescence in the mitochondria. In this way, the degree of opening of mitochondrial MPTP can be judged based on the intensity of the green fluorescence of Calcein in mitochondria. The stronger the green fluorescence, the lower the degree of opening; the weaker the green fluorescence, the higher the degree of opening.

Cell