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MTT Assay

The in vitro determination of toxic effects of u known compounds have been performed by counting viable cells after staining with a vital dye.

 Alternative methods used are measurement of radioisotope incorporation as a measure of DNA synthesis, counting by automated counters and others which rely on dyes and cellular activity. The MTT system is a means of measuring the activity of living cells via mitochondrial dehydrogenases.

The MTT method is simple, accurate and yields reproducible results. The key component is (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) or MTT. Solutions of MTT, dissolved in medium or balanced salt solutions without phenol red, are yellowish in color. Mitochondrial dehydrogenases of viable cells cleave the tetrazolium ring, yielding purple formazan crystals which are insoluble in aqueous solutions. The crystals are dissolved in acidified isopropanol.

The resulting purple solution is spectrophotometrically measured. An increase or decrease in cell number results in a concomitant change in the amount of formazan formed, indicating the degree of cytotoxicity caused by the test material.

     Applications

• Cell Proliferation: effects of cytokines, growth factor, nutrients.
• Cytotoxicity and Apoptosis: evaluation of toxic compounds, anti-cancer antibodies, toxins, environmental pollutants etc.
• Drug Discovery: high-throughput screen for toxic and anticancer drugs.