WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium, monosodium salt), a highly stable and water-soluble WST, is utilized incell Counting Kit-8 (CCK-8).It is more sensitive than WST-1 particularly at neutralpH. Because of the electron mediator, 1-methoxy PMSin this kit is highly stable, and CCK-8 is stable for at least six months at the room temperature and for one year at 0-5°C (asshown in Figure 1).
Advantages: WST-8 is not cell permeable, which results in low cytotoxicity. Therefore,after the assay, it is possible to continue further experiments using the same cells.Furthermore, it produces the water-soluble formazan upon cellular reduction,which would provide an additional advantage to the method by allowing a simpler assay procedure and not required an extra step to dissolve the formazan.
Disadvantages: An important consideration is that reduction of assay substrates isimpacted by changes in intracellular metabolic activity that has no direct effect on over-all cell viability.