mtt（MTT）.doc

mtt（MTT） MTT experiment Principle; Step; Matters needing attention； I. principle The full name of MTT is 3- (4,5) -dimethylthiahiazo (-z-y1) -3,5-di- phenytetrazoliumromide, Chinese chemical name is 3- (4 5-, two -2 -2, 5- methylthiazole) two phenyl tetrazole bromide, trade name: thiazole blue. Its a yellow dye. MTT colorimetric assay is a method of detecting cell survival and growth. The detection principle for live cell mitochondrial succinate dehydrogenase in the exogenous MTT reduced to water insoluble formazan crystal violet (Formazan) and deposited in cells, cell death without this function. Two dimethyl sulfoxide (DMSO) can dissolve formazan in the cells, using enzyme-linked immunosorbent assay to determine the absorbance value at the wavelength of 490nm, can reflect the number of living cells. In the range of a certain number of cells, the amount of MTT crystallization is proportional to the number of cells. The method has been widely used in the detection of some bioactive factors, large-scale screening of antitumor drugs, cytotoxicity tests, and radiosensitivity of tumors. It is characterized by high sensitivity and economy. Disadvantages: as the formazan product MTT by reduction produced insoluble in water, to be dissolved to detect. Not only does this increase the workload, but it also affects the accuracy of the experimental results, and the organic solvent that dissolves one is also harmful to the experimenter. MTT solution preparation: usually, the concentration of MTT in this method is 5mg/ml. Therefore, you can take MTT 0.5 grams, phosphate buffer dissolved in 100 ml (PBS) or phenol red free medium with 0.22 m membrane filter to remove bacteria in solution, 4 degrees can be stored away from light. In the preparation and preservation of the process, the container is best wrapped with aluminum foil. In the experiment, I usually turn off the fluorescent lamp on the ultra clean platform to avoid the light. I think its better. It should be noted that the