A high-throughput fluorimetric assay for angiotensin I-converting enzyme

A high-throughput fluorimetric assay for angiotensin I-converting enzyme

Autor Schwager, Sylva L. Google Scholar
Carmona, Adriana K. Google Scholar
Sturrock, Edward D. Google Scholar
Instituição Univ Cape Town
Universidade Federal de São Paulo (UNIFESP)
Resumo Angiotensin I-converting enzyme (ACE) assays are commonly used for measuring enzymatic activity in clinical and biological samples. the fluorimetric procedure described is sensitive, rapid and involves unsophisticated procedures and inexpensive reagents. It uses the substrate hippuryl-L-histidyl-L-leucine, and the fluorescent adduct of the enzyme-catalyzed product L-histidyl-L-leucine is quantified fluorimetrically. This assay has been adapted for a 96-well plate format that produces comparable data to previously described assays and has the advantage of greater efficiency with respect to both time and reagents. the protocol can be used for routine purposes or for more detailed kinetic analyses. the apparent K(m) and k(cat) values for purified testis ACE determined from a double reciprocal plot were 3.0 mM and 195.7 s(-1), respectively. the protocol can be completed within 4 h.
Idioma Inglês
Data 2006-01-01
Publicado em Nature Protocols. London: Nature Publishing Group, v. 1, n. 4, p. 1961-1964, 2006.
ISSN 1754-2189 (Sherpa/Romeo, fator de impacto)
Editor Nature Publishing Group
Extensão 1961-1964
Fonte http://dx.doi.org/10.1038/nprot.2006.305
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000251155500036
URI http://repositorio.unifesp.br/handle/11600/28655

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