What are the specific protocols used for analyzing MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) in various applications?
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The specific protocols used for analyzing MTT in various applications include the conversion of MTT into purple formazan crystals by living cells, which is then dissolved using a solubilization solution and quantified by measuring the absorbance at 500-600nm using a multi-plate spectrophotometer . Different solvents such as dimethyl sulfoxide, 1% Triton X-100, 1% Bile bovine, 10% sodium dodecyl sulfate, and sodium hydroxide (NaOH) have been evaluated for solubilizing bacterial-formazan, with 1 N NaOH found to be the most effective . The MTT assay has also been used to evaluate the viability of sperm, where MTT is converted into MTT formazan by specific enzymes in viable cells, and the amount of formazan is quantified by measuring absorbance using a spectrophotometer . These protocols provide researchers with a standardized method for assessing cell viability and analyzing the growth of bacterial cells and sperm in various applications.
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24 Citations | The specific protocols used for analyzing MTT in various applications are not mentioned in the provided paper. |
17 Citations | The paper provides a compilation of MTT assay protocols used to determine sperm viability in various animal species and humans, including factors such as sperm and MTT solution preparations, incubation conditions, solubilizing agents, and formazan detection methods. |
20 Citations | The specific protocols used for analyzing MTT in various applications are not mentioned in the provided paper. |
| The specific protocols for analyzing MTT in various applications are not mentioned in the provided paper. The paper is about the MTT assay and its use in characterizing the cytotoxic effects of drugs or nanoparticle formulations on cell cultures. | |
| The specific protocols for analyzing MTT in various applications are not mentioned in the provided paper. The paper is about the MTT assay protocol and the cell lines used in the study. |
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What is the mechanism of the MTT Assay?5 answersThe mechanism of the MTT assay involves the conversion of the tetrazolium salt MTT into insoluble formazan crystals by living cells, specifically through the action of NADPH-dependent oxidoreductase enzymes. Viable cells contain these enzymes, which reduce MTT to formazan. The formazan crystals are then dissolved using a solubilization solution, such as acidified isopropanol or DMSO, resulting in a colored solution. The intensity of the purple color in the solution is directly proportional to the number of viable, metabolically active cells. The absorbance of the colored solution is measured at 500-600nm using a multi-plate spectrophotometer. The MTT assay is widely used to characterize the cytotoxic effects of drugs or nanoparticle formulations on cell lines or primary cell cultures.
What is the MTT assay used for?4 answersThe MTT assay is used to assess cell viability and measure cellular metabolic activity. It is based on the conversion of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) into purple formazan crystals by living cells, indicating their mitochondrial activity. The assay is widely used to evaluate the cytotoxic effects of drugs or nanoparticle formulations on cell lines or primary cell cultures. It is commonly used in studies of cell toxicity and is almost ubiquitous in this field. The MTT assay can be used to quickly and efficiently evaluate the antibacterial activity of natural compounds, making it an important method in natural antibacterial component discovery. However, it is important to optimize the assay and interpret the data carefully to prevent misleading conclusions about variables such as cell viability, treatment toxicity, and cell metabolism.
What role of DMSO in MTT assay?3 answersDMSO is used as a solvent in the MTT assay to dissolve the insoluble formazan crystals. The MTT assay is based on the conversion of MTT into formazan crystals by living cells, which determines the mitochondrial activity of these cells. The formazan crystals are insoluble in water, so DMSO is used to solubilize them and create a colored solution that can be quantified. The use of DMSO as a solvent allows for accurate measurement of the optical densities directly on the MultiScreen plate, improving the reproducibility and accuracy of the assay. DMSO has the same optical refraction index as the filters used in the assay, making it possible to measure the absorbance without loss of cells or formazan crystals.
How to prepare MTS?2 answersThe method for preparing an MT resin involves several steps. In the first step, anhydrous toluene, anhydrous ethanol, and an alkoxyl silane are added to a reaction vessel, followed by the dropwise addition of acetic acid under specific conditions. In the second step, the solvents in the reaction vessel are removed. Finally, in the third step, tetramethyldihydrodisiloxane is added to the reaction vessel, followed by the dropwise addition of acetic acid, resulting in the formation of the MT resin. The molar ratios between the alkoxyl silane and acetic acid, as well as between the tetramethyldihydrodisiloxane and alkoxyl silane, are specified. This method provides a simple and effective way to prepare the MT resin, which exhibits a stable structure.