The effect of in vitro heating on the distribution of nuclear matrix polypeptides in HeLa cells.

Details

Serval ID
serval:BIB_2DA05C4D8248
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
The effect of in vitro heating on the distribution of nuclear matrix polypeptides in HeLa cells.
Journal
Cell Biology International
Author(s)
Martelli A.M., Bareggi R., Riederer B.M., Marugg R.A., Narducci P.
ISSN
1065-6995 (Print)
ISSN-L
1065-6995
Publication state
Published
Issued date
1994
Volume
18
Number
3
Pages
151-158
Language
english
Abstract
The in situ nuclear matrix was obtained from HeLa cells. After permeabilization with nonionic detergent, the resulting structures were incubated for 1 h at 37 degrees C to determine whether or not such an incubation might result in the redistribution of nuclear polypeptides which resisted extraction with buffers of high-ionic strength (1.6 M NaCl or 0.25 M (NH4)2SO4 as well as DNase I digestion. Using indirect immunofluorescence experiments and monoclonal antibodies we show that heating to 37 degrees C changes the distribution of a 160 kDa protein previously shown to be a component of the inner matrix network. On the other hand, a 125 kDa polypeptide was not affected at all by the incubation. Our results clearly indicate that the inclusion of a 37 degrees C incubation (for example during digestion with DNase I) in the protocol to obtain the in situ nuclear matrix can result in the formation of in vitro artifacts.
Keywords
Blotting, Western, Fluorescent Antibody Technique, HeLa Cells/chemistry, Hot Temperature, Humans, Immunohistochemistry, Nuclear Matrix/chemistry, Nuclear Proteins/analysis
Pubmed
Web of science
Create date
24/01/2008 14:34
Last modification date
20/08/2019 13:12
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