This Mitochondrial Permeability Transition Pore Assay is an assay kit for measuring Mitochondrial Permeability Transition Pore activity in cells.
Applications
Functional Studies
Assay Type
Qualitative
Principle of Assay
Cells are loaded with Calcein AM, which is converted intracellularly to fluorescent calcein. Cobalt quenches cytoplasmic fluorescence, leaving mitochondrial fluorescence; opening of the permeability transition pore causes loss of mitochondrial signal.
Platform
Flow Cytometer, Fluorescence Microscope
Sample Type
Cultured cells
Assay Time
60-90 minutes
Components
Calcein AM (1000X), CoCl2 (100X), Ionomycin (200X), Assay Buffer
Storage
Shipped at 4°C. Store at -20°C in the dark. Do not use past expiration date!
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
L929 cells were incubated with Calcein AM (1X), and the cytoplasm including mitochondria emitted strong green fluorescence (A). After cells were further incubated with cobalt chloride (1 mM), the green fluorescence of Calcein in the cytoplasm was quenched by the cobalt chloride, leaving only the green fluorescence in mitochondria (B). Cells treated with ionomycin (0.25 µM) induced a large amount of extracellular Ca2+ to enter the cell. Excessive Ca2+ in the mitochondrial matrix, resulted in the opening of the MPTP, partial release of Calcein from the mitochondria, and the introduction of cobalt ions into the mitochondria and green fluorescence quenching of Calcein (C).
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