Madesh M, Balasubramanian K A
Wellcome Trust Research Laboratory, Department of Gastrointestinal Sciences, Christian Medical College Hospital, Vellore, India.
Indian J Biochem Biophys. 1998 Jun;35(3):184-8.
A simple microtiter plate based colorimetric assay for superoxide dismutase is described. The method, involves generation of superoxide by pyrogallol autoxidation and the inhibition of superoxide dependent reduction of the tetrazolium dye MTT [3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyl tetrazolium bromide] to its formazan, measured at 570 nm. The reaction was terminated by the addition of dimethyl sulfoxide (DMSO) which also helps to solubilize the formazan formed and the colour evolved was stable for many hours. The method was compared with other known methods to measure the activity of purified erythrocyte Cu,ZnSOD and superoxide dismutase activity from various rat tissues. This procedure involves inexpensive reagents, allows a rapid and sensitive measurement of SOD activity and the microtiter plate assay is suitable for use with large number of samples.
描述了一种基于微量滴定板的超氧化物歧化酶比色测定法。该方法包括通过邻苯三酚自氧化产生超氧阴离子，并抑制超氧阴离子依赖的四氮唑染料MTT [3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑]还原为其甲臜，在570 nm处进行测量。通过加入二甲基亚砜（DMSO）终止反应，DMSO也有助于溶解形成的甲臜，并且所产生的颜色在数小时内保持稳定。将该方法与其他已知方法进行比较，以测量纯化的红细胞铜锌超氧化物歧化酶的活性以及来自各种大鼠组织的超氧化物歧化酶活性。该程序使用廉价的试剂，能够快速、灵敏地测量超氧化物歧化酶活性，并且微量滴定板测定法适用于大量样品。
