A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled with Mass Spectrometry

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F21%3A00554565" target="_blank" >RIV/67985823:_____/21:00554565 - isvavai.cz</a>

Výsledek na webu

<a href="https://link.springer.com/protocol/10.1007/978-1-0716-1266-8_28" target="_blank" >https://link.springer.com/protocol/10.1007/978-1-0716-1266-8_28</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/978-1-0716-1266-8_28" target="_blank" >10.1007/978-1-0716-1266-8_28</a>

Jazyk výsledku

angličtina

Název v původním jazyce

A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled with Mass Spectrometry

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled with Mass Spectrometry

Popis výsledku anglicky

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.

Druh

C - Kapitola v odborné knize

CEP obor

—

OECD FORD obor

10609 - Biochemical research methods

Projekt

<a href="/cs/project/NV17-31564A" target="_blank" >NV17-31564A: Aplikace látek pro zefektivnění neinvazivní léčby fibrózního stavu u onemocnění pes equinovarus</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název knihy nebo sborníku

Mitochondrial Medicine

ISBN

978-1-0716-1266-8

Počet stran výsledku

14

Strana od-do

383-396

Počet stran knihy

459

Název nakladatele

Springer

Místo vydání

New York

Kód UT WoS kapitoly

000893349700029