Preparation method of MTT solution
Usually the MTT concentration is 5 mg/ml. Therefore, 0.5 g of MTT can be weighed, dissolved in 100 ml of phosphate buffer (PBS) or phenol red free medium, filtered through a 0.22 μm filter to remove bacteria from the solution, and stored at 4 ° C in the dark. . The container is preferably wrapped in aluminum foil during the preparation and storage process.
It should be noted that the MTT method can only be used to detect the relative number and relative viability of cells, but not the absolute number of cells. In order to ensure the linearity of the experimental results when using the microplate reader, the MTT absorbance is preferably in the range of 0-0.7.
MTT is generally best used now, filtered at 4 ° C in the dark for 2 weeks, or 5mg / ml stored at -20 degrees for long-term preservation, to avoid repeated freezing and thawing, preferably small doses, with light Bags or black paper, tin foil paper to protect from light to avoid decomposition. I usually put the MTT powder in the EP tube. When it is used, it is added directly to the culture plate. There is no need to match it at all, especially when the MTT turns grayish green.
MTT is carcinogenic and should be used with care. It is best to have a transparent film glove. The formulated MTT needs to be sterile, MTT is very sensitive to bacteria; it does not matter if it is not protected from light when it is added to the 96-well plate. After all, the time is short, or you can turn off the illumination on the console when you are not at ease. When preparing MTT, it was dissolved in PBS, and it was also formulated with physiological saline and dissolved in a 60 ° C water bath.
PBS preparation method: Nacl 8g, Kcl 0.2g, Na2HPO4 1.44g, KH2PO4 0.24g, adjusted to pH 7.4, to a volume of 1L.
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