|Product name||Mitochondrial Permeability Transition Pore Assay|
|Applications notes||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.|
|Kit components||• Calcein AM (1000×)
• CoCl2 (100×)
• Ionomycin (200×)
• Assay Buffer
|Features & Benefits||• High safety: almost no toxicity to cells;
• Compatibility with multiple detection methods: can be used in flow cytometry and fluorescence microscopy.
|Usage notes||• Do not mix or substitute reagents or materials from other kit lots or vendors.|
|Storage instructions||Stored at -20℃ for 12 months, protected from light|
|Shipping||Gel pack with blue ice.|
|Precautions||The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.|
|Background||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.|
Fig. L929 cells was incubated with L929 cells Calcein AM (1X), and the cytoplasm including mitochondria emitted strong green fluorescence (A). After cells were further incubated with CoCl2 (1mM), the green fluorescence of Calcein in the cytoplasm was quenched by CoCl2, leaving only the green fluorescence in mitochondria (B). Cells treated with Ionomycin (0.25μM) induced a large amount of extracellular Ca2+ into the cell, excessive Ca2+ into the mitochondrial matrix, resulting in the opening of 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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1.The species of antibody reactivity should be the sample species that can be matched normally after Abbkine R&D experts have passed strict scientific verification. If your sample is not within the range of reactivity, in order to improve the efficiency and results of your experiment, it is not suggested to try other species. Otherwise, it may lead to sample mismatch and affect the effect of your experiment.
2.Please aliquot the antibody received as soon as possible and store it at -20℃, avoid repeated freezing and thawing, and use it within one year.
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