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[求助]
關(guān)于NO測定和計算問題
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因?yàn)槭卟私M織本身含有亞硝酸鹽,無法使用griess試劑法,采用血紅蛋白分析法測定蔬菜組織NO,方法如下: NO accumulation was assayed and calculated by following the conversion of oxyhemoglobin (HbO2) to methemoglobin (MetHb) spectrophotometrically at 401 nm and 421 nm, using an extinction coefficient of 77 mm−1 cm−1 (A401 HbO2, A421 MetHb). the samples were homogenized in a mortar with 100 mM K-phosphate buffer (pH 7.0) and 0.6% (w:v) insoluble polyvinylpolypyrrolidone. The extract was clarified by adding powdered activated carbon and centrifuged at 11 000 × g for 10 min at 4 ◦C. The supernatant was filtered through a PTFE Millipore membrane (0.45 m) and immediately assayed for NO. Five minutes before oxyhemoglobin addition, samples were pretreated with catalase (100 U) and superoxide dismutase (100 U) to remove ROS. 問題來了,這個怎么計算結(jié)果的? |
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