Examination and analyses to the coloring reacting of MTT
ZHAO Cheng-yan
JING Zhi-an
NIU Qing-xia
Abstract：Objective: To improve experimental conditions of the colorimetric assay, depends on the reduction by living cells oftetrazolium salt, MTT, and to enhance sensitivity and reliability and to lower background. Methods: The eolorimetric assay withMTT, and the experiments of cytotoxicity and proliferation of cells. Results: The analyses and examining to the experimentalconditions of the colorimetric assay with MTT have found that majority of the mitochondrial enzyme succinate-dehydrogenase isdistributed over surface of cell membrane; the enviromment of reaction of enzyme with MTT is pH 6.7; following cells incubatedin the connon medium containing serums and phenol red were completed, the supematant is discarded, and leaving cells alonereact with MTr, and subsequent use of pure propanol to rapidly dissolve the formazan. The pure organic solvent can precipitatetrace proteins to be removed by centrifugation and lower acid organic solvent (pH5.8) can also eliminate to the influence ofphenol red to the assay. Cotclusion: When the experimental conditions have be improved, the cure of cell enzyme reactionwith MTT and the cure of MTT cleaved reaction-time under in saturated state are linear relationship. The background is verylowed down;the sensitivity and reliability are heightened;the minimum detectable number of K-562 and Human T-cells are respectively 1000 cells and 10000 cells in our hands.
Keywords：MTTenzyme release waycolorimetric assay
Publication Date：2000-01-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：6( 107-111,116 )