Gel electrophoretic quantitation of protein carbonyls derivatized with tritiated sodium borohydride

Liang-Jun Yan, Rajindar S. Sohal

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

A method for the quantitation of protein carbonyls, which have been widely employed as markers of protein oxidative damage, is described. Protein carbonyls were derivatized with tritiated sodium borohydride and the tritiated proteins were separated on SDS-PAGE. Protein bands, visualized by Coomassie blue staining, were then excised and incubated in 30% H2O2 at 60°C for 48 h. Tritium, incorporated into the proteins, was quantitated by liquid scintillation counting after gel solubilization by H2O2. This method can be applied to the measurement of carbonylation of specific proteins as it employs SDS-PAGE and has the advantage that unreacted NaB3H4 in the labeling reaction mixture need not be removed. The present method, when combined with immunochemical detection of protein carbonyls, should be very useful in the quantitation of oxidative damage to individual proteins.

Original languageEnglish
Pages (from-to)176-182
Number of pages7
JournalAnalytical Biochemistry
Volume265
Issue number1
DOIs
StatePublished - 1 Dec 1998

Keywords

  • Aging
  • Gel electrophoresis
  • Oxidative damage
  • Protein carbonyls
  • Tritiated sodium borohydride

Fingerprint Dive into the research topics of 'Gel electrophoretic quantitation of protein carbonyls derivatized with tritiated sodium borohydride'. Together they form a unique fingerprint.

  • Cite this