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Mitochondrial Medicine
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SYSNO ASEP 0554565 Document Type M - Monograph Chapter R&D Document Type Monograph Chapter Title A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled with Mass Spectrometry Author(s) Kohutiar, M. (CZ)
Eckhardt, Adam (FGU-C) RID, ORCIDSource Title Mitochondrial Medicine, Assessing Mitochondria, 2. - New York : Springer, 2021 / Weissig Volkmar ; Edeas Marvin - ISSN 1064-3745 - ISBN 978-1-0716-1266-8 Pages s. 383-396 Number of pages 14 s. Number of pages 459 Publication form Online - E Language eng - English Country US - United States Keywords mitochondria ; nitrotyrosine ; 2D electrophoresis ; immunoblotting ; mass spectrometry OECD category Biochemical research methods R&D Projects NV17-31564A GA MZd - Ministry of Health (MZ) Institutional support FGU-C - RVO:67985823 UT WOS 000893349700029 EID SCOPUS 85107188199 DOI https://doi.org/10.1007/978-1-0716-1266-8_28 Annotation Nitrotyrosine formation is caused by presence of reactive oxygen and nitrogen species. Nitration is a very selective process leading to specific modification of only a few tyrosines in protein molecule. 2D electrophoresis and western blotting techniques coupled with mass spectrometry are common methods used in analysis of proteome. Here we describe protocol for analysis of peroxynitrite-induced protein nitration in isolated mitochondria. Mitochondrial proteins are separated by 2D electrophoresis and transferred to nitrocellulose membrane. Membranes are then incubated with antibodies against nitrotyrosine. Positive spots are compared with corresponding Coomassie-stained gels, and protein nitration is confirmed with mass spectrometry techniques. Workplace Institute of Physiology Contact Lucie Trajhanová, lucie.trajhanova@fgu.cas.cz, Tel.: 241 062 400 Year of Publishing 2022 Electronic address https://link.springer.com/protocol/10.1007/978-1-0716-1266-8_28
Number of the records: 1