Mitochondrial Permeability Transition Pore Assay

Mitochondrial permeablity transition pore (MPTP), also known as magachannel, is a non selective high conductivity channel that exists between the inner and outer membranes of mitochondria and is composed of multiple protein complexes. MPTP may be involved in the release of mitochondrial components during cell death. The normal mitochondrial inner membrane of a cell 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 transition pore switches back and forth between opening and closing. During apoptosis and pathological death of cells, the permeability of mitochondrial membrane potential transfer pore changes, Ca2+ overload, mitochondrial glutathione oxidation, increase of reactive oxygen species, including subsequent release of cytochrome C, and mitochondrial membrane potential decline all lead to the activation of MPTP. Mitochondrial Permeability Transition Pore Assay Kit s a more direct detection method for detecting the opening of MPTP than only based on mitochondrial membrane potential analysis.

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• High safety: almost no toxicity to cells; • Compatibility with multiple detection methods: can be used in flow cytometry and fluorescence microscopy.

Mitochondrial Permeability Transition Pore Assay Kit is a more direct detection method for detecting the opening of MPTP than only based on mitochondrial membrane potential analysis. The principle is: Firstly, loading Calcein AM through passive transportation, which is a kind of cell staining reagent for fluorescent labeling of 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 CoCl2, the fluorescence from the cytoplasm is quenched by CoCl2, only left fluorescence in the mitochondria. As a control, cells can be loaded with Calcein AM and CoCl2, and treated with Ionomycin, so as to make cells load more Ca2+, which causes the activation of MPTP and quenching of mitochondrial fluorescence.