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. 1988 Sep 1;254(2):519–523. doi: 10.1042/bj2540519

Free-radical generation by copper ions and hydrogen peroxide. Stimulation by Hepes buffer.

J A Simpson 1, K H Cheeseman 1, S E Smith 1, R T Dean 1
PMCID: PMC1135108  PMID: 3178771

Abstract

Hydroxyl radicals (OH.), generated by a phosphate-buffered Cu2+/H2O2 system, were detected by lucigenin-amplified chemiluminescence, deoxyribose degradation and benzoate hydroxylation. In each system the buffer, Hepes, was found to stimulate radical generation significantly. There are two main reasons for this effect: Hepes increases Cu2+ solubility in phosphate-buffered systems, and forms a complex with Cu2+ that is effective in generating OH. from H2O2. Pipes, a structurally similar buffer, and histidine, a known Cu2+ chelator, were found to have a similar effect. These data suggest that the crucial factor in such free-radical-generating systems is the availability of Cu2+, and that these actions of Hepes should be considered in the design of studies utilizing such systems.

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Selected References

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