Lj. Yan et Rs. Sohal, Gel electrophoretic quantitation of protein carbonyls derivatized with tritiated sodium borohydride, ANALYT BIOC, 265(1), 1998, pp. 176-182
A method for the quantitation of protein carbonyls, which have been widely
employed as markers of protein oxidative damage, is described. Protein carb
onyls 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 degrees C
for 48 h. Tritium, incorporated into the proteins, was quantitated by Liqu
id 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 (NaBH4)-H-3 in the la
belling reaction mixture need not be removed. The present method, when comb
ined with immunochemical detection of protein carbonyls, should be very use
ful in the quantitation of oxidative damage to individual proteins. (C) 199
8 Academic Press.