| The TACS MTT Cell Proliferation
Assay (MTT-CPA) is a sensitive kit for the measurement of cell
proliferation based upon the reduction of the tetrazolium salt
3,[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT).
Changes in cell proliferative activity caused by trophic factors, growth
inhibitors, or inducers and inhibitors of apoptosis, may be quantified
using the MTT-CPA. MTT is reduced to an insoluble formazan dye by
mitochondrial enzymes associated with metabolic activity. The reduction
of MTT is primarily due to glycolytic activity within the cell and is
dependent upon the presence of NADH and NADPH.
Common methods for determining cell viability depend upon membrane
integrity (e.g. trypan blue exclusion), or incorporation of nucleotides
during cell proliferation (e.g. BrdU or 3H-thymidine). These methods are
limited by the impracticality of processing large numbers of samples, or
by the requirement for handling hazardous materials. The MTT Assay, in
contrast, provides a rapid and versatile method for assessing cell
viability.
The assay is used to measure changes in cell proliferation. In
actively proliferating cells, an increase in MTT conversion is
spectrophotometrically quantified. Comparison of this value to an
untreated control provides a relative increase in cellular proliferative
activity. Conversely, in cells that are undergoing apoptosis, MTT
reduction decreases, reflecting the loss of cell viability. |
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