Mitochondrial Permeability Transition Pore Assay Kit or MPTP Assay Kit, 100-1000 T

Brands:Beyotime

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Place of Origin:China

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Shipping:Blue Ice/Dry Ice

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Component:Assay Buffer, Calcein AM (1000X), CoCl2 (100X), Ionomycin (200X), Solubilizer (100X)

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Shelf Life:12 months

Storage:-20℃

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Em:517nm

Ex:494nm 

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Test Principle:Beyotime's MPTP Assay Kit employs the acetoxymethyl ester of calcein dye, calcein AM, which passively diffuses into cells and accumulates in cytosolic compartments, including the mitochondria. Once inside cells, intracellular esterases cleave the acetoxymethyl esters to liberate the very polar fluorescent dye calcein. The Calcein exhibiting bright green fluorescence does not cross the membrane and retains in mitochondria and cytosol in appreciable amounts over relatively short periods. Under physical pH conditions, the fluorescence of calcein can be quenched by metal ions, such as Co2+、Ni2+、Cu2+、Fe3+, and Mn2+. In this assay, the fluorescence from cytosolic calcein is quenched by the addition of CoCl2, while the fluorescence from the mitochondrial calcein is maintained because MPTP is closed in healthy cells and CoCl2 could not enter into mitochondria. As a control, cells that have been loaded with calcein AM and CoCl2 can also be treated with ionomycin to allow entry of excess Ca2+ into the cells to trigger the opening of MPTP and subsequent loss of mitochondrial calcein fluorescence. In this way, the opening of MPTP can be evaluated based on the intensity of green fluorescence in mitochondria. The stronger the green fluorescence, the less opened MPTP. The weaker the green fluorescence, the more opened MPTP.

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Detection Instrument :fluorescence microscope, flow cytometer, or fluorescence microplate reader

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