[問題] 抗氧化酵素POD跟SOD的測定
POD 的活性藉由測量維他命C在H2O2的氧化程度。
加入 50μl 懸浮物到950 μl potassium phosphate buffer (pH 7.0, 50 mM)(含0.1
mM EDTA、 0.5 mM ascorbate、and 0.1 mM H2O2。)後,維他命C開始氧化。反應30秒後
,以分光光度計在波長290 nm時測量到吸收減少。在20°C進行分析。 One unit of POD
was defined as the variance of absorbance per 107 cells (ΔA470/107 cells)。
問題一 請問維他命C氧化程度愈高 是否表POD愈沒用?
二 維他命C氧化程度愈高 顏色會愈淺 吸收愈少嗎?
測量SOD 的活性, the reaction buffer (700μl) was prepared with 50 mM
potassium phosphate buffer (pH 7.8), 0.1 mM EDTA, 18 μM cytochrome c, and
0.1 mM xanthine.
加黃飄呤氧化脢(Xanthine oxidase)
到reaction buffer to get an increase of absorbance at 550 nm of ca. 0.025 min
-1 at 25°C.
Then, 15 to 100μl of supernatant sample and H2O was added to the reaction
buffer, including xanthine oxidase, to give a volume of 800 μl.
The rate of cytochrome c was
measured at 550 nm for an additional minute.
One unit of SOD was defined as the variance of absorbance per min per 107
cells ( A550/min 107 cells)。
問題三 請問SOD Xanthine oxidase cytochrome c 這三者間關係為何阿??
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※ 編輯: mgc2 來自: 210.209.177.21 (10/02 20:11)
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