DCFH-DA体外测材料ROS问题？ 2.6. chemical ros assayto study the insonation induced ros generation in solution, rb4was tested using a modified 2′, 7′–dichlorofluorescin diacetate (dcfhda) assay. dcfh-da (1 ml, 1 mm in meoh, aldrich) was hydrolyzed innaoh aqueous solution (0.01 n, 4 ml) at rt for 30 min to yield a nonfluorescent dcfh intermediate. the solution was neutralized with20 ml of nah2po4 (25 mm) and shielded with aluminum foil. the finalsolution of dcfh was around 40 μm. rb4 (1.0 mg) was dissolved indmso (1 ml), and then diluted with water into a 20 μm solution. therb4 solution (20 μm, 10 ml) was mixed with a dcfh solution (40 μm,10 ml) as the test solution. the test solution (0.5 ml) was placed intoeach well of a 24-well plate. the dcfh solution without rb4 wasincluded as a background control. the plates were treated withultrasound (0.4–1.2 watt/cm2) one well by one well for 30 s; theinsonated wells were then checked using a fluorescence plate reader,ex 485 nm/em 520 nm， 查看更多
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