2026-08-16T14:15:21Zhttps://docta.ucm.es/rest/oai/request
oai:docta.ucm.es:20.500.14352/945982024-06-14T23:47:32Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense author Linares Gómez, María author Marín-García, Patricia author Méndez, Darío author Puyet Catalina, Antonio author Díez Martín, Amalia author Bautista Santa Cruz, José Manuel 2024-01-23T08:49:25Z 2024-01-23T08:49:25Z 2011 J. Proteome Res. 2011, 10, 4, 1719–1727Publication Date:January 15, 2011 https://doi.org/10.1021/pr101014e 1535-38931535-3907 10.1021/pr101014e https://hdl.handle.net/20.500.14352/94598 https://doi.org/10.1021/pr101014e Oxidative stress plays a critical role in the pathogenesis of a number of diseases. The carbonyl end products of protein oxidation are among the most commonly measured markers of oxidation in biological samples. Protein carbonyl functional groups may be derivatized with 2,4-dinitrophenylhydrazine (DNPH) to render a stable 2,4-dinitrophenylhydrazone-protein (DNP-protein) and the carbonyl contents of individual proteins then determined by two-dimensional electrophoresis followed by immunoblotting using specific anti-DNP antibodies. Unfortunately, derivatization of proteins with DNPH could affect their mass spectrometry (MS) identification. This problem can be overcome using nontreated samples for protein identification. Nevertheless, derivatization could also affect their mobility, which might be solved by performing the derivatization step after the initial electrophoresis. Here, we compare two-dimensional redox proteome maps of mouse cerebellum acquired by performing the DNPH derivatization step before or after electrophoresis and detect differences in protein patterns. When the same approach is used for protein detection and identification, both methods were found to be useful to identify carbonylated proteins. However, whereas pre-DNPH derivatized proteins were successfully analyzed, high background staining complicated the analysis when the DNPH reaction was performed after transblotting. Comparative data on protein identification using both methods are provided. eng Attribution-NonCommercial-NoDerivatives 4.0 International Proteomic Approaches to Identifying Carbonylated Proteins in Brain Tissue journal article 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 URL https://docta.ucm.es/bitstreams/7e2744b8-5fca-4dc9-8eaf-ad7a23be26eb/download File MD5 59ca4e4af88fd63d990c8b70e33df10b 5139309 application/pdf Proteomic Approaches _version aceptada.pdf