Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells
A thymidine analog, 5-ethynyl-2′-deoxyuridine (EdU), has been reported as a rapid labeling tool for direct measurement of cells in S-phase. The alkynyl group of EdU is a biologically inert group that undergoes an extremely selective reaction with azido-functionalized groups via Cu(I)-catalyzed alkyn...
| Автори: | , , , |
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| Формат: | Стаття |
| Мова: | English |
| Опубліковано: |
Taylor & Francis Group
2010-07-01
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| Серія: | BioTechniques |
| Предмети: | |
| Онлайн доступ: | https://www.future-science.com/doi/10.2144/000113463 |
| _version_ | 1827008022060204032 |
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| author | Kai Li L. Andrew Lee Xiaobing Lu Qian Wang |
| author_facet | Kai Li L. Andrew Lee Xiaobing Lu Qian Wang |
| author_sort | Kai Li |
| collection | DOAJ |
| description | A thymidine analog, 5-ethynyl-2′-deoxyuridine (EdU), has been reported as a rapid labeling tool for direct measurement of cells in S-phase. The alkynyl group of EdU is a biologically inert group that undergoes an extremely selective reaction with azido-functionalized groups via Cu(I)-catalyzed alkyneazide cycloaddition (CuAAC or “click”) reaction. Here we report the highly efficient reaction of the terminal alkynyl group of EdU with a pro-fluorogenic compound, 3-azido-7-hydroxycoumarin, to afford an intense fluorescent 1,2,3-triazole product, which occurs only after the CuAAC reaction. This new method eliminates concerns for residual fluorescence since the unreacted precursors are optically inactive. The procedure therefore does not require extensive wash steps to remove the unreacted fluorescent dyes in the sample, allowing for immediate quantification and visualization after the reaction. The advantage over currently available commercial products is its potential to streamline high-throughput applications and help minimize errors. |
| first_indexed | 2025-02-18T12:27:57Z |
| format | Article |
| id | doaj.art-08b51c87965c4474b1661143d3e915aa |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2025-02-18T12:27:57Z |
| publishDate | 2010-07-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-08b51c87965c4474b1661143d3e915aa2024-11-02T04:16:40ZengTaylor & Francis GroupBioTechniques0736-62051940-98182010-07-0149152552710.2144/000113463Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cellsKai Li0L. Andrew Lee1Xiaobing Lu2Qian Wang31State Key Laboratory of Fine Chemicals, Dalian University of Technology (DUT), Dalian, China2Department of Chemistry and Biochemistry and Nanocenter, University of South Carolina, Columbia, SC, USA1State Key Laboratory of Fine Chemicals, Dalian University of Technology (DUT), Dalian, China2Department of Chemistry and Biochemistry and Nanocenter, University of South Carolina, Columbia, SC, USAA thymidine analog, 5-ethynyl-2′-deoxyuridine (EdU), has been reported as a rapid labeling tool for direct measurement of cells in S-phase. The alkynyl group of EdU is a biologically inert group that undergoes an extremely selective reaction with azido-functionalized groups via Cu(I)-catalyzed alkyneazide cycloaddition (CuAAC or “click”) reaction. Here we report the highly efficient reaction of the terminal alkynyl group of EdU with a pro-fluorogenic compound, 3-azido-7-hydroxycoumarin, to afford an intense fluorescent 1,2,3-triazole product, which occurs only after the CuAAC reaction. This new method eliminates concerns for residual fluorescence since the unreacted precursors are optically inactive. The procedure therefore does not require extensive wash steps to remove the unreacted fluorescent dyes in the sample, allowing for immediate quantification and visualization after the reaction. The advantage over currently available commercial products is its potential to streamline high-throughput applications and help minimize errors.https://www.future-science.com/doi/10.2144/0001134635-ethynyl-2′-deoxyuridinecell proliferationfluorogenic reactionCu-catalyzed alkyne-azide cycloadditionclick chemistry |
| spellingShingle | Kai Li L. Andrew Lee Xiaobing Lu Qian Wang Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells BioTechniques 5-ethynyl-2′-deoxyuridine cell proliferation fluorogenic reaction Cu-catalyzed alkyne-azide cycloaddition click chemistry |
| title | Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells |
| title_full | Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells |
| title_fullStr | Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells |
| title_full_unstemmed | Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells |
| title_short | Fluorogenic “click” reaction for labeling and detection of DNA in proliferating cells |
| title_sort | fluorogenic click reaction for labeling and detection of dna in proliferating cells |
| topic | 5-ethynyl-2′-deoxyuridine cell proliferation fluorogenic reaction Cu-catalyzed alkyne-azide cycloaddition click chemistry |
| url | https://www.future-science.com/doi/10.2144/000113463 |
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