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小木虫论坛-学术科研互动平台 » 生物医药区 » 动植物 » 综合其他 » 测植物叶片硝酸还原酶活性的问题
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[求助] 测植物叶片硝酸还原酶活性的问题

各位大侠，我现在正在测植物叶片硝酸还原酶活性，所用的方法是活体法，NR activity was assayed in vivo by measuring NO-2 production in tissue that has been vacuum infiltrated with buffered NO-3 solutions.[31]The leaves and roots from the nonstressed and stressed seedlings were collected in the morning between 8:30 and 11:00 solar time. Circles 5mm in diameter were cut from the leaves of O. europaea. Leaves and roots of R. sphaerocarpa and roots of O. europaea were cut into 5mm sections.
Approximately 300 mg of leaf punches and 300 mg of roots from each
plant were placed into tubes containing 2mL of an incubation medium
consisting of 0.05M tris-HCl pH 7.8 and 0.25M KNO3. The tubes
were sealed and kept in the dark at 30C during 1 h. The nitrite released
into the medium was determined after incubation by treating 1mL of
the aliquots with 1mL of 1% sulphanilamide in 1M HCl and 1mL of
0.01% N-1-napthyl-ethylenediamine hydrochloride (NNEDA). After
15 min, the optical density was measured at 540nm with Beckman
spectrophotometer.
上面的这些是我在一篇SCI 文献里看到的，老师要求按这方法做，但是其中的试剂配制方法，参比溶液的设置，还有标准曲线的绘制，我都没有，而且我查了很多文献都没有具体的操作步骤。希望各位高人能够帮小弟一把！给我一些上面方法的具体操作步骤，不胜感激。[ Last edited by amisking on 2011-6-15 at 19:38 ]
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