Abstract：
:We have developed a photometric, platereader-based microassay for superoxide anion scavening activity in vitro. Superoxide anions were generated using a xanthine oxidase/hypoxanthine system and detected by following the reduction of ferricytochrome c at 550 nM. Inhibitory activity was assessed for superoxide dismutase (SOD) and the superoxide anion scavengers tiron and TEMPO together with a number of TEMPO derivatives. The initial rate of change in optical density (OD) at 550 nm, i.e., initial reaction rate, generated by xanthine oxidase (20 mU/ml)/hypoxanthine (100 μM) coupled to ferricytochrome c (100 μM) was effectively abolished by SOD (200 U/ml), tiron (10 mM) and TEMPO (0.3 mM), indicating the involvement of superoxide anions. TEMPO derivatives inhibited the initial reaction rate with the potency order: TEMPO > 4-hydroxy-TEMPO = 4-carboxy-TEMPO. In contrast, 4-hydroxy-TEMPO, which lacks the free radical nitroxide function, was inactive up to 1 mM.
journal_name
Environ Toxicol Pharmacoljournal_title
Environmental toxicology and pharmacologyauthors
Laight DW,Andrews TJ,Haj-Yehia AI,Carrier MJ,Anggård EEdoi
10.1016/s1382-6689(96)00143-3subject
Has Abstractpub_date
1997-02-15 00:00:00pages
65-8issue
1eissn
1382-6689issn
1872-7077pii
S1382-6689(96)00143-3journal_volume
3pub_type
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