Complex I spectrophotometric assay in cultured cells: Detailed analysis of key factors
Data
2013-04-01
Autores
Oliveira, Katia Klug 
Kiyomoto, Beatriz Hitomi
Rodrigues, Andresa De Santi
Tengan, Celia Harumi
Tipo
Artigo
Título da Revista
ISSN da Revista
Título de Volume
Resumo
The diagnosis of mitochondrial encephalomyopathies caused by complex I (C-I) deficiency relies mainly on the spectrophotometric C-I assay. Considered difficult, this assay lacks reliability and has high nonspecific activity. We studied the key factors of this assay in cultured cells (cybrid and fibroblast): ubiquinone analogues, rotenone inhibition to determine specific activity, and mode of permeabilization of mitochondria] membranes. We showed that ubiquinone 1 allows a more stable and reliable assay, better results were obtained with rotenone inhibition done in the same assay, and mitochondria] permeability was improved just by freeze/thawing the sample prior to the assay. (C) 2012 Elsevier Inc. All rights reserved.
Descrição
Citação
Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 435, n. 1, p. 57-59, 2013.