Cell Cycle Analysis Using In Vivo Staining of DNA-Synthesizing Cells
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
31079341
DOI
10.1007/7651_2019_228
Knihovny.cz E-resources
- Keywords
- Antibodies, Cell cycle, DNA labellin`g, Flow cytometry, Immunophenotyping, S-phase,
- MeSH
- Data Analysis MeSH
- Staining and Labeling methods MeSH
- Bromodeoxyuridine metabolism MeSH
- Cell Differentiation MeSH
- Cell Cycle * MeSH
- Bone Marrow Cells metabolism MeSH
- Deoxyuridine analogs & derivatives MeSH
- DNA biosynthesis MeSH
- Immunophenotyping MeSH
- Mitosis MeSH
- Mice MeSH
- Flow Cytometry MeSH
- Rheology MeSH
- S Phase MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 5-ethynyl-2'-deoxyuridine MeSH Browser
- Bromodeoxyuridine MeSH
- Deoxyuridine MeSH
- DNA MeSH
The thymidine analogues BrdU (5-bromo-2´-deoxyuridine) and EdU (5-ethynyl-2´-deoxyuridine) are routinely used for determination of the cells synthesizing DNA in the S-phase of the cell cycle. Availability of the anti-BrdU antibody clone MoBu-1 detecting only BrdU allowed to develop a method for the sequential DNA labelling by these two thymidine analogues for determining the cell cycle kinetic parameters.In the current step-by-step protocol, we present` two approaches optimized for in vivo study of the cell cycle and the limitations that such approaches imply: (1) determination of the cell flow rate into the G2-phase by dual EdU/BrdU DNA-labelling method and (2) determination of the outflow of DNA-labelled cells arising from the mitosis.
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