Two-dimensional gel electrophoretic detection of protein carbonyls derivatized with biotin-hydrazide

Jinzi Wu, Xiaoting Luo, Siqun Jing, Liang Jun Yan

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Protein carbonyls are protein oxidation products that are often used to measure the magnitude of protein oxidative damage induced by reactive oxygen or reactive nitrogen species. Protein carbonyls have been found to be elevated during aging and in age-related diseases such as stroke, diabetes, and neurodegenerative diseases. In the present article, we provide detailed protocols for detection of mitochondrial protein carbonyls labeled with biotin-hydrazide followed by 2-dimensional isoelectric focusing (IEF)/SDS-PAGE and Western blotting probed with horse-radish peroxidase-conjugated streptavidin. The presented procedures can also be modified for detection of carbonylation of non-mitochondrial proteins.

Original languageEnglish
Pages (from-to)128-131
Number of pages4
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume1019
DOIs
StatePublished - 15 Apr 2016

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Keywords

  • Biotin-hydrazide
  • Carbonylation
  • IEF/SDS-PAGE
  • Mitochondria
  • Protein carbonyls
  • Protocol
  • Streptavidin

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