Two-dimensional gel electrophoretic detection of protein carbonyls derivatized with biotin-hydrazide

Jinzi Wu; Xiaoting Luo; Siqun Jing; Liang Jun Yan

doi:10.1016/j.jchromb.2015.11.003

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 journal › Article › peer-review

8 Scopus citations

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 language	English
Pages (from-to)	128-131
Number of pages	4
Journal	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume	1019
DOIs	https://doi.org/10.1016/j.jchromb.2015.11.003
State	Published - 15 Apr 2016

Keywords

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

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10.1016/j.jchromb.2015.11.003

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title = "Two-dimensional gel electrophoretic detection of protein carbonyls derivatized with biotin-hydrazide",

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.",

keywords = "Biotin-hydrazide, Carbonylation, IEF/SDS-PAGE, Mitochondria, Protein carbonyls, Protocol, Streptavidin",

author = "Jinzi Wu and Xiaoting Luo and Siqun Jing and Yan, {Liang Jun}",

note = "Funding Information: LJY was supported in part by the National Institutes of Health (Grant: R01NS079792). Publisher Copyright: {\textcopyright} 2015 Elsevier B.V.",

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doi = "10.1016/j.jchromb.2015.11.003",

language = "English",

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journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",

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TY - JOUR

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

N2 - 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.

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.

KW - Biotin-hydrazide

KW - Carbonylation

KW - IEF/SDS-PAGE

KW - Mitochondria

KW - Protein carbonyls

KW - Protocol

KW - Streptavidin

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U2 - 10.1016/j.jchromb.2015.11.003

DO - 10.1016/j.jchromb.2015.11.003

M3 - Article

C2 - 26590475

AN - SCOPUS:84978319130

SN - 1570-0232

VL - 1019

SP - 128

EP - 131

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

ER -

Two-dimensional gel electrophoretic detection of protein carbonyls derivatized with biotin-hydrazide

Abstract

Keywords

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