Mitochondria are the energy centers of cells and play a very important role in the processes of apoptosis and necrotic cell death. Mitochondrial permeability transition pore (mPTP), also known as mitochondrial megachannel (MMC), It is a non-specific channel composed of the inner and outer membranes of mitochondria, which participates in the release of substances within mitochondria during cell death. The opening of mitochondrial permeability transition pores is a key event causing cell death and plays an important role in the regulation of cell survival and apoptosis. The detection principle of this kit: The kit uses Calcein AM to stain cells, causing the cytoplasm, including mitochondria, to show strong green fluorescence. Under physiological pH conditions, when Calcein complexes with Co2+, the fluorescence signal is quenched. Under normal circumstances, the mPTP in mitochondria is off, and at this time,Co2+ cannot enter the mitochondria, resulting in only green fluorescence appearing within the mitochondria. The inner mitochondrial membrane of normal cells 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 conversion pore switches back and forth between opening and closing. When cells undergo apoptosis and pathological death, the permeability of mitochondrial membrane potential conversion pores changes. Overload of Ca2+, oxidation of mitochondrial glutathione, increase in reactive oxygen species levels, including subsequent release of cytochrome C, and decrease in mitochondrial membrane potential can all lead to the activation of mitochondrial permeability conversion pores. Further treatment with the calcium ion carrier Ionomycin as a control to induce a large amount of extracellular Ca2+ to enter the intracellular and mitochondrial matrix and cause the opening of mPTP will weaken or disappear the green fluorescence in the mitochondria. In this way, the degree of opening of mitochondrial MPTP can be judged based on the intensity of the green fluorescence of Calcein in mitochondria. The stronger the green fluorescence, the lower the degree of opening; the weaker the green fluorescence, the higher the degree of opening.
The fluorescent probe provided in this kit specifically targets mitochondria in living cells, which are oxidized by mitochondrial superoxides to produce bright red fluorescence. The probe excited at 396 nm can specifically detect superoxide of supermitochondria, and the mitochondrial reactive oxygen species (ROS) can also be detected using the excitation wavelength of 510 nm.
Mitochondrial stress refers to the stress state that occurs when mitochondria are damaged or overloaded under the influence of external or internal factors (such as oxidative stress, nutritional deficiency, toxins, etc.). This kind of stress can affect the energy production of mitochondria, leading to insufficient cellular energy and possibly triggering cell damage, inflammatory responses or promoting cell apoptosis. Long-term mitochondrial stress is closely related to various diseases (such as neurodegenerative diseases, cardiovascular diseases, metabolic diseases, etc.) and the aging process. This kit provides an oxygen-sensitive fluorescent probe, which increases the fluorescence of the probe by the cells consuming oxygen. After adding the regulators oligomycin, carboxycyanide-4-trifluoromethoxylhydrazone (FCCP) and antimycin A, the effect of the metabolic process on maximum respiration can be visually presented. The addition of oligomycin blocked ATP synthesis, slowed the electron transfer rate, reduced the respiratory rate and decreased oxygen consumption. FCCP is used to study the respiratory activity of mitochondria in the uncoupled state. After the uncoupled oxidative phosphorylation of FCCP, the proton pump of the electron transport chain (ETC) is no longer limited by ATP synthesis, the electron transport rate increases, and the oxygen consumption significantly increases, which is used to determine the maximum respiratory capacity. Antimycin A inhibits the main respiratory chain, and the electron transfer dependent on complex III stops, resulting in a significant decrease in basal oxygen consumption, which is used to measure the reserve respiratory rate.
Monoacylglycerol lipase (MAGL) is a widely distributed serine hydrolase. MAGL can not only decompose triacylglycerol, which plays an important role in lipid metabolism, but also hydrolyze 2-arachidonic acid and regulate the signaling of the cannabinoid system in the body. The principle of this kit is that the substrate is catalyzed by MAGL, and the product generated can react with the chromogenic agent. By measuring the amount of chromogenic substance produced per unit time, the MAGL enzyme activity in the sample can be calculated.
Monoamine Oxidase (MAO) is a mitochondrial membrane binding enzyme, which is widely present in connective tissues and cell mitochondria of liver, kidney, small intestine, stomach, brain and other organs. MAO-A metabolizes neurotransmitters such as serotonin, dopamine, serotonin, epinephrine and norepinephrine. MOA inhibition increases levels of neurotransmitters and are used to treat depression, neurodegenerative diseases, and cardiovascular disease. The detection principle of this kit: MAO-A can catalyze the substrate to produce fluorescent substances. After adding inhibitors, the generation of fluorescent substances will be inhibited, and the effect of inhibitors will be determined according to the degree of inhibition.
MAO can catalyze 4-dimethylambenzylamine to produce p-dimethylaminobenzaldehyde. p-Dimethylaminobenzaldehyde has a characteristic absorption peak at 355 nm. The activity of MAO can be calculated indirectly by analyzing the production of p-dimethylaminobenzaldehyde.
MAO can catalysis 4-dimethylambenzylamine to produce p-dimethylaminobenzaldehyde. p-Dimethylaminobenzaldehyde has a characteristic absorption peak at 355 nm. The activity of MAO can be calculated indirectly by analyzing the production of p-dimethylaminobenzaldehyde.
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