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Microassay of Superoxide Anion Scavenging Activity in Vitro

Microassay of Superoxide Anion Scavenging Activity in Vitro

D W Laight, T J Andrews, A I Haj-Yehia, M J Carrier, E E Anggård

Environ Toxicol Pharmacol. 1997 Feb 15;3(1):65-8.

PMID: 21781760

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP37149181	4-Carboxy-TEMPO, free radical		37149-18-1	Price

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