Mitochondrial Permeability Transition Pore (MPTP) Assay Kit

1. Calcein AM (1000X) decomposes easily in humid environments. It is recommended to aliquot it appropriately and store it at -20°C in a dry， sealed container. 2. Fluorescent probe is easily to quench， avoid light during storage and usage. 3. CoCl? is corrosive and harmful to humans. Wear protective gear during handling to avoid inhalation or direct skin contact. CoCl? must not be released directly into the environment. 4. For your safety and health， wear a lab coat and disposable gloves during operation. 5. This product is for research use only. It must not be used for clinical diagnosis or treatment. Mitochondria serve as the cellular powerhouses， playing vital roles in numerous cellular activities. The Mitochondrial Permeability Transition Pore (MPTP) is a channel on mitochondria composed of both the inner and outer mitochondrial membranes. It participates in the release of mitochondrial contents during cell death processes such as apoptosis. Consequently， the opening of MPTP is considered a key indicator of cell death mechanisms like apoptosis. The Mitochondrial Permeability Transition Pore Assay Kit is a kit that detects the degree of opening of the mitochondrial permeability transition pore using the fluorescent probe calcein AM. Calcein AM is a fluorescent probe capable of labeling live cells. It readily enters cells and is cleaved by intracellular esterases into green-fluorescent calcein， which remains within the cell. CoCl? binds to calcein and quenches its fluorescence. However， under normal conditions， the mitochondrial MPTP remains closed， preserving the green fluorescence of calcein within the mitochondria. However， when the calcium ionophore ionomycin induces extracellular Ca2? influx into the cell， opening the MPTP， mitochondrial calcein binds with CoCl?， leading to further or complete fluorescence quenching. Thus， the intensity of green fluorescence within mitochondria indicates the degree of MPTP opening: stronger green fluorescence corresponds to lower MPTP opening. If the decoupling agent CCCP is used instead of the calcium ionophore ionomycin， the mitochondrial membrane potential is lost but MPTP remains closed for a short time， resulting in no change in mitochondrial fluorescence. CCCP is not provided with this kit; purchase it separately using the product number if needed. This kit provides 100 assays for flow cytometry or 1000 assays for 96-well plate detection.