The fingerprint of anti-bromodeoxyuridine antibodies and its use for the assessment of their affinity to 5-bromo-2'-deoxyuridine in cellular DNA under various conditions

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15110%2F15%3A33154080" target="_blank" >RIV/61989592:15110/15:33154080 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388963:_____/15:00446574

Výsledek na webu

<a href="http://www.plosone.org/article/fetchObject.action?uri=info:doi/10.1371/journal.pone.0132393&representation=PDF" target="_blank" >http://www.plosone.org/article/fetchObject.action?uri=info:doi/10.1371/journal.pone.0132393&representation=PDF</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1371/journal.pone.0132393" target="_blank" >10.1371/journal.pone.0132393</a>

Jazyk výsledku

angličtina

Název v původním jazyce

The fingerprint of anti-bromodeoxyuridine antibodies and its use for the assessment of their affinity to 5-bromo-2'-deoxyuridine in cellular DNA under various conditions

Popis výsledku v původním jazyce

We have developed a simple system for the analysis of the affinity of anti-bromodeoxyuridine antibodies. The system is based on the anchored oligonucleotides containing 5-bromo-2'-deoxyuridine (BrdU) at three different positions. It allows a reliable estimation of the reactivity of particular clones of monoclonal anti-bromodeoxyuridine antibodies with BrdU in fixed and permeabilized cells. Using oligonucleotide probes and four different protocols for the detection of BrdU incorporated in cellular DNA, we identified two antibody clones that evinced sufficient reactivity to BrdU in all the tested protocols. One of these clones exhibited higher reactivity to 5-iodo-2'-deoxyuridine (IdU) than to BrdU. It allowed us to increase the sensitivity of the used protocols without a negative effect on the cell physiology as the cytotoxicity of IdU was comparable with BrdU and negligible when compared to 5-ethynyl-2'-deoxyuridine. The combination of IdU and the improved protocol for oxidative degrad

Název v anglickém jazyce

The fingerprint of anti-bromodeoxyuridine antibodies and its use for the assessment of their affinity to 5-bromo-2'-deoxyuridine in cellular DNA under various conditions

Popis výsledku anglicky

We have developed a simple system for the analysis of the affinity of anti-bromodeoxyuridine antibodies. The system is based on the anchored oligonucleotides containing 5-bromo-2'-deoxyuridine (BrdU) at three different positions. It allows a reliable estimation of the reactivity of particular clones of monoclonal anti-bromodeoxyuridine antibodies with BrdU in fixed and permeabilized cells. Using oligonucleotide probes and four different protocols for the detection of BrdU incorporated in cellular DNA, we identified two antibody clones that evinced sufficient reactivity to BrdU in all the tested protocols. One of these clones exhibited higher reactivity to 5-iodo-2'-deoxyuridine (IdU) than to BrdU. It allowed us to increase the sensitivity of the used protocols without a negative effect on the cell physiology as the cytotoxicity of IdU was comparable with BrdU and negligible when compared to 5-ethynyl-2'-deoxyuridine. The combination of IdU and the improved protocol for oxidative degrad

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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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)

Rok uplatnění

2015

Kód důvěrnosti údajů

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

Název periodika

PLoS One

ISSN

1932-6203

e-ISSN

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Svazek periodika

10

Číslo periodika v rámci svazku

7

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

16

Strana od-do

"e0132393-1"-"e0132393-16"

Kód UT WoS článku

000358162300128

EID výsledku v databázi Scopus

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