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 Citations (Scopus)

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

Fingerprint

Gels
Proteins
Neurodegenerative diseases
Raphanus
Carbonylation
Reactive Nitrogen Species
Streptavidin
Mitochondrial Proteins
Isoelectric Focusing
Medical problems
Neurodegenerative Diseases
Peroxidase
Horses
Polyacrylamide Gel Electrophoresis
Aging of materials
Western Blotting
Stroke
biotin hydrazide
Oxygen
Oxidation

Keywords

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

Cite this

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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.",
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T1 - Two-dimensional gel electrophoretic detection of protein carbonyls derivatized with biotin-hydrazide

AU - Wu, Jinzi

AU - Luo, Xiaoting

AU - Jing, Siqun

AU - Yan, Liang-Jun

PY - 2016/4/15

Y1 - 2016/4/15

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AB - 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.

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KW - Carbonylation

KW - IEF/SDS-PAGE

KW - Mitochondria

KW - Protein carbonyls

KW - Protocol

KW - Streptavidin

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