Ethanol dehydrogenase can catalyze oxidative dehydrogenation of ethanol to acetaldehyde, and NAD+ is reduced to produce NADH. NADH, under the action of 1-mPMS, transfer electrons to WST-8 to produce the yellow product, which has a characteristic absorption peak at 450 nm. Therefore, ethanol content can be quantified by measure the OD value at 450 nm.
The extracellular acidification rate (ECAR) refers to the effect of acidic substances released by cells during metabolism on the extracellular environment, which is an important indicator to evaluate the energy metabolism and growth status of cells. During cellular metabolism, energy generation requires either aerobic or anaerobic respiration. When cells undergo anaerobic respiration, a large number of acidic substances are produced, which are pumped by the cell membrane into the extracellular environment, leading to the acidification of the extracellular environment. Therefore, ECAR is an important indicator to evaluate the acidic substances produced by anaerobic respiration during cell metabolism, and its change can reflect the degree of cell metabolism. The kit uses a specific probe (Ex/Em: 490 nm/535 nm) that changes the fluorescence value when the pH of the extracellular environment changes. The acidified extracellular environment will reduce the fluorescence value of the probe to detect the extracellular acidification rate.
Fatty acid amide hydrolase (FAAH) is a membrane binding protein belonging to the serine hydrolase family. FAAH hydrolyzes many important endogenous fatty acid amides, including endocannabinoid (AEA) and palmitoyl glycolamide (PEA). FAAH plays a key role in controlling cannabinoid signaling by hydrolyzing 2arachidenol glycerol in AEA and certain tissues. Detection principle: FAAH specifically cut the substrate, the fluorophores emit fluorescence, and detect fluorescence at the excitation wavelength of 340 nm and the emission wavelength of 440 nm. Because FAAH is a serine protein, FAAHspecific inhibitors are used in the kit to inhibit enzyme activity and exclude the effects of non-specific enzyme activity.
Fatty acid oxidation (FAO) is the main pathway of fatty acid decomposition in the body, which can supply a large amount of energy required by the organism. FAO is also the transformation process of fatty acids. The length of fatty acid chain required by the organism is different. Through oxidation, long-chain fatty acids can be transformed into fatty acids with suitable growth degree for the organism metabolism. The detection principle of this kit: FAO process consumes the substrate and NAD+, and generates NADH. Under the action of electron coupling agent and chromogenesis agent, the orange red substance is generated, which can be detected at 450 nm.
Fatty acid is an important source of energy for the body. Fatty acid intake capacity has been associated with diseases such as obesity and diabetes, and is therefore an important target for the treatment of diseases such as obesity, Ⅱ diabetes, and fatty liver. Hyperactive cancer cells have a strong fatty acid uptake capacity, so the fatty acid uptake capacity is also used as an important indicator of cancer drug development, and the fatty acid pathway is also attracted attention as one of the targets of cancer drug development. This kit provides a convenient fluorescence assay to measure the cell's ability to take up fatty acids. Fatty acid substrate can enter the cell through the fatty acid transporter on the cell membrane, and the ability of cells to take up fatty acid can be detected by fluorescence microscopy, fluorescence microplate reader, flow cytometry and other instruments to detect the fluorescence intensity.
Ferroptosis inhibitory protein-1(FSP-1) induce non-caspase-dependent apoptosis based on factors such as C-terminal fragment of amino acid series, nuclear translocation and overexpression. Through the FSP1-CoQ10-NAD(P)H pathway and the classical glutathione (GSH)-GPX4 pathway, FSP-1 can protect cells from ferroptosis, and plays a "double-edged sword" role in cell life activities. The detection principle of this kit: FSP-1 catalyzes the substrate reaction to generate NADH, so that its fluorescence value increases under the excitation wavelength of 535 nm and emission wavelength of 587 nm. Adding inhibitors will inhibit the activity of FSP-1 and cause the rate of increase in fluorescence value to decrease, and the activity of FSP-1 can be calculated by measuring the difference value.
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