Protein carbonylation detection methods: A comparison
Esra'a Alomari,
Stefano Bruno,
Luca Ronda,
Gianluca Paredi,
Stefano Bettati,
Andrea Mozzarelli
Affiliations
Esra'a Alomari
Department of Food and Drug, University of Parma, Parma, Italy
Stefano Bruno
Department of Food and Drug, University of Parma, Parma, Italy; Biopharmanet-TEC, University of Parma, Parma, Italy; Corresponding author at: Department of Food and Drug, University of Parma Parma, Italy.
Luca Ronda
Department of Medicine and Surgery, University of Parma, Parma, Italy; Biopharmanet-TEC, University of Parma, Parma, Italy
Gianluca Paredi
Department of Food and Drug, University of Parma, Parma, Italy
Stefano Bettati
Department of Medicine and Surgery, University of Parma, Parma, Italy; Biopharmanet-TEC, University of Parma, Parma, Italy; Istituto Nazionale Biostrutture e Biosistemi, Rome, Italy
Andrea Mozzarelli
Department of Food and Drug, University of Parma, Parma, Italy; Biopharmanet-TEC, University of Parma, Parma, Italy; Istituto Nazionale Biostrutture e Biosistemi, Rome, Italy; Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Pisa, Italy
The data reported here are a comparison among four different methods for the detection of carbonylated proteins, a validated biomarker of oxidative stress. The reference samples were heart and kidney extracts of Guinea pigs transfused with hemoglobin-based oxygen carriers (Alomari et al. FRBM, [11]). We measured the carbonyl content of organ extracts by using i) the Levine spectrophotometric method, which takes advantage of the chromogenic reaction of carbonyl groups with 2,4-dinitrophenylhydrazine (DNPH), ii) a commercially available ELISA assay based on an anti-DNPH antibodies, iii) a commercially available Western blot method based on anti-DNPH antibodies and iv) an in-gel detection approach with the fluorophoric reagent fluorescein-5-thiosemicarbazide. The former two methods measure total protein carbonylation of a sample, whereas the latter two require an electrophoretic separation and therefore potentially allow for the identification of specific carbonylated proteins.
