Document Detail


Multiparameter cell cycle analysis.
MedLine Citation:
PMID:  21116986     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cell cycle-related cytometry and analysis is an essential experimental paradigm for the cell biology of yeast, mammalian, and drosophila cells. Methods have not changed much for many years. The most common is DNA content analysis, which has been well-published and reviewed. Next most common is analysis of 5-bromo-2-deoxyuridine (BrdU) incorporation, detected by specific antibodies - also well-published and reviewed. A new measurement approach to S phase labeling utilizes 5'-ethynyl-2'-deoxyuridine (EdU) incorporation and a chemical reaction to label substituted DNA. The approach is new, but published work indicates that it is equivalent to BrdU incorporation. Finally, multiple antibody labeling to detect epitopes on cell cycle-regulated proteins is the most complex of the cytometric cell cycle assays, requiring knowledge of the chemistry of fixation, the biochemistry of antibody-antigen reactions, and spectral compensation. Because all of this knowledge is relatively well presented, methodologically, in many papers and reviews, this chapter presents a bare-bones Methods section for one mammalian cell type and an extended Notes section, focusing on aspects that are problematic or not well described in the literature.
Authors:
James W Jacobberger; R Michael Sramkoski; Tammy Stefan
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  699     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2011  
Date Detail:
Created Date:  2010-11-30     Completed Date:  2011-05-23     Revised Date:  2013-02-11    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  229-49     Citation Subset:  IM    
Affiliation:
Cytometry and Imaging Microscopy Core, Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, USA.
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MeSH Terms
Descriptor/Qualifier:
Antibodies / metabolism
Cell Cycle*
Cell Line, Tumor
Data Collection
Data Display
Flow Cytometry / methods*
Fluorescent Dyes / metabolism
Humans
Staining and Labeling
Grant Support
ID/Acronym/Agency:
P30CA43703/CA/NCI NIH HHS; R01CA73413/CA/NCI NIH HHS; UL1 RR024989/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies; 0/Fluorescent Dyes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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