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| rdf:type
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| Description
| - 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 have performed 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 with fluorescent 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 suppression of EdU reactivity with anti-BrdU antibodies: a method based on hydrochloric acid treatment and a method based on the use of copper(I) ions. The use of hydrochloric acid resulted in significant increase of the non-specific signal. In the case of the second method, no such effect was observed.
- 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 have performed 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 with fluorescent 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 suppression of EdU reactivity with anti-BrdU antibodies: a method based on hydrochloric acid treatment and a method based on the use of copper(I) ions. The use of hydrochloric acid resulted in significant increase of the non-specific signal. In the case of the second method, no such effect was observed. (en)
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| Title
| - Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine — The Method for the Effective Suppression of This Cross-Reactivity
- Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine — The Method for the Effective Suppression of This Cross-Reactivity (en)
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| skos:prefLabel
| - Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine — The Method for the Effective Suppression of This Cross-Reactivity
- Most Anti-BrdU Antibodies React with 2-Deoxy-5-Ethynyluridine — The Method for the Effective Suppression of This Cross-Reactivity (en)
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| skos:notation
| - RIV/68081707:_____/12:00386136!RIV13-AV0-68081707
|
| http://linked.open...avai/riv/aktivita
| |
| http://linked.open...avai/riv/aktivity
| - P(GBP302/12/G157), P(KAN200520801), Z(AV0Z40550506), Z(AV0Z50040702), Z(AV0Z50520514)
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| http://linked.open...iv/cisloPeriodika
| |
| http://linked.open...vai/riv/dodaniDat
| |
| http://linked.open...aciTvurceVysledku
| |
| http://linked.open.../riv/druhVysledku
| |
| http://linked.open...iv/duvernostUdaju
| |
| http://linked.open...titaPredkladatele
| |
| http://linked.open...dnocenehoVysledku
| |
| http://linked.open...ai/riv/idVysledku
| - RIV/68081707:_____/12:00386136
|
| http://linked.open...riv/jazykVysledku
| |
| http://linked.open.../riv/klicovaSlova
| - 2´-deoxy-5-ethynyluridine; 5-bromo-2´-deoxyuridine; DNA replication; anti-BrdU antibodies; immunocytochemistry (en)
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| http://linked.open.../riv/klicoveSlovo
| |
| http://linked.open...odStatuVydavatele
| - US - Spojené státy americké
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| http://linked.open...ontrolniKodProRIV
| |
| http://linked.open...i/riv/nazevZdroje
| |
| http://linked.open...in/vavai/riv/obor
| |
| http://linked.open...ichTvurcuVysledku
| |
| http://linked.open...cetTvurcuVysledku
| |
| http://linked.open...vavai/riv/projekt
| |
| http://linked.open...UplatneniVysledku
| |
| http://linked.open...v/svazekPeriodika
| |
| http://linked.open...iv/tvurceVysledku
| - Rosenberg, Ivan
- Koberna, Karel
- Ligasová, Anna
- Liboska, Radek
- Strunin, Dmytro
|
| http://linked.open...ain/vavai/riv/wos
| |
| http://linked.open...n/vavai/riv/zamer
| |
| issn
| |
| number of pages
| |
| http://bibframe.org/vocab/doi
| - 10.1371/journal.pone.0051679
|
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