Document Detail

FACS-based detection of phosphorylated histone H3 for the quantitation of mitotic cells.
MedLine Citation:
PMID:  15220538     Owner:  NLM     Status:  MEDLINE    
The G2 checkpoint blocks cells from entering mitosis when DNA is damaged, and helps to protect the integrity of the genome. Tumor cells contain mutations that can inactivate checkpoints, and the inactivation of the G2 checkpoint can induce genomic instability and alter cellular responses to chemotherapeutic agents that damage DNA. The traditional method to assess whether the G2 checkpoint is normal is to microscopically count mitotic cells. A method using the fluorescence-activated cell scanner (FACS) is described, based on the presence of an intra-nuclear antigen present only in mitotic cells, detected using a specific, commercially available antibody. Cell staining and FACS analysis can be done in a single day, making this a rapid and reliable method to quantitate mitotic cells for various applications.
William R Taylor
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  281     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2004  
Date Detail:
Created Date:  2004-06-28     Completed Date:  2004-09-16     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  293-9     Citation Subset:  IM    
Department of Biological Sciences, University of Toledo, OH, USA.
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MeSH Terms
Fibrosarcoma / chemistry,  metabolism
Flow Cytometry / methods*
Fluorescein / diagnostic use
G2 Phase*
Histones / metabolism*
Serine / chemistry
Tumor Cells, Cultured
Grant Support
Reg. No./Substance:
0/Histones; 2321-07-5/Fluorescein; 56-45-1/Serine

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