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NBT assay
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| Principle: |
| ˇ@ˇ@Superoxide anion was generated by xanthine-xanthine oxidase and detected by the NBT reduction. NBT is a reagent that can absorb superoxide and change it color to purple(absorbed at 570nm). We can use this character to detect it OD value every 20 seconds in 20 min. Thus, we can get the slope of rate that NBT binds to superoxide anion. |
| Meterials & Methods: |
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ˇ@ˇ@The reaction mixture in a final volume of
1 ml contained potassium phosphate buffer(50mM,pH7.4),Na2EDTA(15mM), hypoxanthine(3mM
in 50mM KOH), NBT(0.6mM) and xanthine oxidase(1 unit in 10ml buffer). |