Format

Send to

Choose Destination
Chem Res Toxicol. 1993 Jul-Aug;6(4):430-3.

Immunochemical detection of oxidized proteins.

Author information

1
Occupational and Environmental Health Program, University of Arkansas for Medical Sciences, Little Rock 72205-7199.

Abstract

An immunochemical assay was developed to detect carbonyl moieties that result from oxidative damage to proteins. Bovine serum albumin was reacted with hydroxyl radicals generated via a Fenton-like mechanism or by a radiolysis mechanism. The resulting albumin-derived carbonyls were reacted with 2,4-dinitrophenylhydrazine, giving the corresponding hydrazones, which were detected by Western blot using anti-dinitrophenyl antisera. The immunoblot demonstrated a concentration-dependent increase in carbonyl formation, as well as fragmentation of the albumin into two distinct bands with molecular masses of 51 and 45 kDa when oxidized with the Fenton-like mechanism, and 62 and 46 kDa when oxidized by radiolysis. Analysis of the immunoblot using laser densitometry indicated a linear relationship between carbonyl groups and increasing treatment from radiolysis. This immunochemical assay was approximately 3 orders of magnitude more sensitive than the spectrophotometric method and was able to determine the molecular mass of carbonyl-modified polypeptides in the detection of oxidative damage.

PMID:
8374038
DOI:
10.1021/tx00034a007
[Indexed for MEDLINE]

Supplemental Content

Loading ...
Support Center