Hi, I want to do MTT assay with my cell model, which is related mitochondrial. Somebody suggests me to use XTT assay instead of MTT. What is difference between XTT and MTT?
Thank you for your help.
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Yes, I think it is not good choice if MTT assay in invovle mitochondria. XTT may be better than MTT. But XTT is expensive.
Both dyes look at redox potential. Major difference is that MTT forms an insoluble formazan derivative on oxidation (requiring you to permeabilize and solubilize with DMSO/acid-alcohol etc), whereas XTT forms a soluble colored product. There are various reports on sensitivity, but it probably depends on cell type/application/media that you use. As Robertwe said, XTT is more expensive than MTT.
You can also try using any of the fluorescent redox dyes sold by Sigma or Mol Probes if you want to study mitochondrial function (DCFDA, JC1 etc).
MTT is a colorimetric assay that measures the reduction of yellow MTT by mitochondrial succinate
dehydrogenase. The MTT enters the cells and passes into the mitochondria where it is
reduced to an insoluble, coloured (dark purple) formazan product. The cells are then
solubilised with an organic solvent (eg. isopropanol) and the released, solubilised
formazan reagent is measured spectrophotometrically. Since reduction of MTT can only
occur in metabolically active cells the level of activity is a measure of the viability of the
HI... I want to know how MTT enters the cell and act with mitochondrial enzymes.