Journal
ANALYTICAL BIOCHEMISTRY
Volume 458, Issue -, Pages 69-71Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2014.04.034
Keywords
Oxidative stress; Protein oxidation; Carbonyl quantification; DNPH assay
Funding
- Fundacao para a Ciencia e Tecnologia (FCT, Portugal)
- FEDER (European Fund for Regional Development)-COMPETE-QREN-EU [PEst-C/QUI/UI0686/2011 (FCOMP-01-0124-FEDER-022 716)]
- FCT [SFRH/BD/64189/2009]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/64189/2009] Funding Source: FCT
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This work proposes a modification of the 2,4-dinitrophenylhydrazine (DNPH) spectrophotometric assay commonly used to evaluate the concentration of carbonyl groups in oxidized proteins. In this approach NaOH is added to the protein solution after the addition of DNPH, shifting the maximum absorbance wavelength of the derivatized protein from 370 to 450 nm. This reduces the interference of DNPH and allows the direct quantification in the sample solution without the need for the precipitation, washing, and resuspension steps that are carried out in the traditional DNPH method. The two methods were compared under various conditions and are statistically equivalent. (C) 2014 Elsevier Inc. All rights reserved.
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