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SOD活性的测定采用邻苯三酚自氧化法。 (1)邻苯三酚自氧化速率的测定:在20 mL试管中加入9 mL50 mmol/L Tris-HCl缓冲液,于25℃恒温水浴中放置20 min,用微量注射器加入40 µL25℃预热的45 mmol/L邻苯三酚溶液(用10 mmol/L HCl配制,空白管加Tris-HCl缓冲液),立即混匀,准确反应3 min后,用微量注射器迅速加入50 µL5 %Vc溶液,立即混匀,倒入3 cm光径的比色杯中,每隔30 s测定A420一次。计算线性范围内每分钟A420的增值,此即为邻苯三酚的自氧化速率。 (2)SOD活力的测定:加入邻苯三酚前,先加一定体积的溶液,蒸馏水减少相应体积,其它条件均与上述一致,计算加酶后邻苯三酚的自氧化速率。 (3)SOD活力计算方法: 式中,A0——自氧化速率; As——加入粗酶液后的氧化速率; V ——加样体积(mL); N ——样品稀释倍数。 |
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wangwei2008
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本人不才,帮你试着翻译一下,不当之处,还望见谅! SOD activity was determined by pyrogallol autoxidation method. (1) Determinationof pyrogallol auto-oxidation rate : added 9 mL 50 mmol / L Tris-HCl buffer In a 20 mL test tube, at 25 ℃ water bath for 20 min, added 40 µL25℃ prewarmed 45 mmol / L pyrogallol solution (preparation by 10 mmol /L HCl , a blank tube plus Tris-HCl buffer) by micro-syringe, mixed immediately and after reacted accurately to 3 min, quickly adding 50 µL5 %Vc solution by micro-syringe, mixed immediately, poured into 3cm optical path colorimetric cup, measured at A420 once every 30s . Calculated within the linear range of value-added at A420 which is the pyrogallol self-oxidation rate per minute. (2) Determination of SOD activity : Before adding pyrogallol, added a certain volume of solution, a corresponding reduction in volume of distilled water, and other conditions are in line with the above, calculated pyrogallol self-oxidation rate after adding enzyme. (3) SOD activity calculated as follows: 式中,A0——自氧化速率; As——加入粗酶液后的氧化速率; V ——加样体积(mL); N ——样品稀释倍数。 Where, A0 - self-oxidation rate; As - the oxidation rate after adding the crude enzyme solution; V - loading volume (mL); N - time of dilution of sample. |
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