Staining of Proliferating Cell Nuclei with Biotinylated dUTP
The bulletin of the Yamaguchi Medical School Volume 38 Issue 3-4
Page 41-48
published_at 1991-12
Title
Staining of Proliferating Cell Nuclei with Biotinylated dUTP
Creators
Murakami Tomoyuki
Source Identifiers
Creator Keywords
Biotinylated dUTP
BrdU
DNA systhesis
Flow cytometry
A method of in situ biotinylated dUTP incorporation into nuclei was developed for evaluaton of cell growth activity. HeLa cells or frozen sections of cancer tissues were incubated in a solution containing bio-dUTP and the other deoxyribonucleotides (dATP, dCTP, dGTP), after permeabilization of cell membrane with Triton X-100, and signals from bio-dUTP in nuclei were detected with an avidin-biotin-peroxidase complex (ABC method), or with fluoresceinavidin. This staining procedure was simple and rapid as compared with that for BrdU incorporated into DNA. In HeLa cells, bio-dUTP-incorporated cells could be stained by the ABC method and the labeling index was compatible with that of BrdU, but the intranuclear distribution patterns of bio-dUTP-incorporated cells was also possible. In some frozen sections of cancer tissues, cells synthesizing DNA were labeled with bio-dUTP. These results suggest that this will be a useful method for evaluating cell growth activity, although the biological significance may be different the BrdU incorporation experiment.
Languages
eng
Resource Type
departmental bulletin paper
Publishers
Yamaguchi University Graduate School of Medicine
Date Issued
1991-12
File Version
Version of Record
Access Rights
open access
Relations
[ISSN]0513-1812
[NCID]AA00594272
Schools
医学部