Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine ? The Method for the Effective Suppression of This Cross-Reactivity
Identifikátory výsledku
Kód výsledku v IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F12%3A00386136" target="_blank" >RIV/61388963:_____/12:00386136 - isvavai.cz</a>
Nalezeny alternativní kódy
RIV/68081707:_____/12:00386136 RIV/68378050:_____/12:00386136
Výsledek na webu
<a href="http://dx.doi.org/10.1371/journal.pone.0051679" target="_blank" >http://dx.doi.org/10.1371/journal.pone.0051679</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1371/journal.pone.0051679" target="_blank" >10.1371/journal.pone.0051679</a>
Alternativní jazyky
Jazyk výsledku
angličtina
Název v původním jazyce
Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine ? The Method for the Effective Suppression of This Cross-Reactivity
Popis výsledku v původním jazyce
5-Bromo-2´-deoxyuridine (BrdU) and 2´-deoxy-5-ethynyluridine (EdU) are widely used as markers of replicated DNA. While BrdU is detected by means of antibodies, the click reaction typically with fluorescent azido-dyes is used for EdU localisation. We haveperformed an analysis of ten samples of antibodies against BrdU with respect to their reactivity with EdU. Except for one sample all the others evinced reactivity with EdU. A high level of EdU persists in nuclear DNA even after the reaction of EdU withfluorescent azido-dyes if the common concentration of dye is used. Although a ten-time increase of azido-dye concentration resulted in a decrease of the signal provided by anti-BrdU antibodies, it also resulted in the substantial increase of the non-specific signal. We have shown that this unwanted reactivity is effectively suppressed by nonfluorescent azido molecules. We tested two protocols of BrdU revelation in experiments of the simultaneous localisation of BrdU and EdU with the supp
Název v anglickém jazyce
Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine ? The Method for the Effective Suppression of This Cross-Reactivity
Popis výsledku anglicky
5-Bromo-2´-deoxyuridine (BrdU) and 2´-deoxy-5-ethynyluridine (EdU) are widely used as markers of replicated DNA. While BrdU is detected by means of antibodies, the click reaction typically with fluorescent azido-dyes is used for EdU localisation. We haveperformed an analysis of ten samples of antibodies against BrdU with respect to their reactivity with EdU. Except for one sample all the others evinced reactivity with EdU. A high level of EdU persists in nuclear DNA even after the reaction of EdU withfluorescent azido-dyes if the common concentration of dye is used. Although a ten-time increase of azido-dye concentration resulted in a decrease of the signal provided by anti-BrdU antibodies, it also resulted in the substantial increase of the non-specific signal. We have shown that this unwanted reactivity is effectively suppressed by nonfluorescent azido molecules. We tested two protocols of BrdU revelation in experiments of the simultaneous localisation of BrdU and EdU with the supp
Klasifikace
Druh
J<sub>x</sub> - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)
CEP obor
CC - Organická chemie
OECD FORD obor
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Návaznosti výsledku
Projekt
Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.
Návaznosti
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)
Ostatní
Rok uplatnění
2012
Kód důvěrnosti údajů
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Údaje specifické pro druh výsledku
Název periodika
PLoS ONE
ISSN
1932-6203
e-ISSN
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Svazek periodika
7
Číslo periodika v rámci svazku
12
Stát vydavatele periodika
US - Spojené státy americké
Počet stran výsledku
10
Strana od-do
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Kód UT WoS článku
000312483300023
EID výsledku v databázi Scopus
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