小中大
你说的那种方法我没见过,文献上基本都是用DCF这个荧光染料的，可以用荧光分光光度计,也可以用流式检测。
DCF assay for reactive oxygen species (ROS)
The cellular ROS was quantified as described by Hong and Joseph, 1999. Accumulation of intracellular ROS can be detected by using DCFH-DA, which crosses cell membranes and is hydrolyzed enzymatically by intracellular esterases to nonfluorescent DCFH. In the presence of ROS, DCFH is oxidized to highly fluorescent dichlorofluorescein (DCF), which is readily detected by a fluorescent microplate reader. The cells on poly-L-lysine coated 96-well plates were incubated with 100 AM DCFHDA in the loading medium in 5%CO2/95% air at 37jC for 30 min. After DCFH-DA was removed, the cells were washed once with DMEM and incubated in DMEM containing different concentrations of
scutellarin (0.1–10 AM) or 10 AM vitamin E for 30 min, followed by the addition of 200 AM H2O2. After 30 min incubation in 5%CO2/95% air at 37jC, the fluorescence from each well was captured, digitized, and stored on a computer using 1420 Victor2 V (Perkin Elmer Life Science, USA). The percentage increase in fluorescence per well was calculated by the formula [(Ft30-Ft0)/Ft0 100], where Ft0 is the fluorescence at time 0 min and Ft30 is the fluorescence at time 30 min in the presence of H2O2.
