Immunochemical detection of oxidized proteins
| Autor: | Keller Rj, Jack A. Hinson, Neil R. Pumford, Halmes Nc |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Free Radicals
Radical Blotting Western Toxicology Catalysis Western blot Rosaniline Dyes medicine Fragmentation (cell biology) Bovine serum albumin Chromatography medicine.diagnostic_test biology Molecular mass Chemistry Immunochemistry Albumin Proteins Serum Albumin Bovine Hydrogen Peroxide General Medicine Orders of magnitude (mass) Phenylhydrazines Biochemistry Gamma Rays Immunoglobulin G Radiolysis biology.protein Electrophoresis Polyacrylamide Gel Spectrophotometry Ultraviolet Vanadates Oxidation-Reduction Densitometry |
| Zdroj: | Chemical Research in Toxicology. 6:430-433 |
| ISSN: | 1520-5010 0893-228X |
| DOI: | 10.1021/tx00034a007 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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