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A fluorescence-based protocol for quantifying angiotensin-converting enzyme activity

Nature Protocols volume 1, pages 2423–2427 (2006)Cite this article

Abstract

The determination of angiotensin-converting enzyme (ACE) activity represents a useful tool in the study of different health pathologies, such as hypertension. This protocol describes a fluorescent assay for measuring ACE activity in vitro with high precision and sensitivity. The method relies on the ability of ACE to hydrolyse the internally quenched fluorescent substrate o-aminobenzoylglycyl-p-nitro-L-phenylalanyl-L-proline. The generation of the fluorescent product o-aminobenzoylglycine can be continuously monitored, preferably using a microtiter-plate fluorometer, though the use of a conventional cuvette fluorometer would also be possible. The method has important advantages with respect to other assays, because it involves only a one-step reagent, is easy to carry out and allows the analysis of an elevated number of samples in shorter times. It can be completed in one and a half hours. In addition, the fact that all reagents are commercially available allows the rapid introduction of the assay into the laboratory.

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Acknowledgements

This work was supported by an I3P contract from the European Social Fund (M.A.S.) and by a Marie Curie ERG grant from the European Commission (M.A.S.).

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  1. Instituto de Agroquímica y Tecnología de Alimentos (CSIC), P.O. Box 73, Burjassot (Valencia), 46100, Spain

    Miguel Ángel Sentandreu & Fidel Toldrá

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  1. Miguel Ángel Sentandreu
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  2. Fidel Toldrá
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Correspondence to Miguel Ángel Sentandreu or Fidel Toldrá.

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Ángel Sentandreu, M., Toldrá, F. A fluorescence-based protocol for quantifying angiotensin-converting enzyme activity. Nat Protoc 1, 2423–2427 (2006). https://doi.org/10.1038/nprot.2006.349

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