EurekaMag.com logo
+ Site Statistics
References:
52,725,316
Abstracts:
28,411,598
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Preparation of nuclear matrices from cultured cells subfractionation of nuclei in situ


Journal of Cell Biology 98(5): 1886-1894
Preparation of nuclear matrices from cultured cells subfractionation of nuclei in situ
Analyses of the different structural systems of the nucleus and the proteins associated with them pose many problems. Because these systems are largely overlapping, in situ localization studies that preserve the in vivo location of proteins and cellular structures often are not satisfactory. Biochemical cell fractionation may provide artifactual reuslts due to cross-contamination of extracts and structures. A method was developed that combines biochemical cell fractionation and in situ localization and leads to the preparation of a residual cellular skeleton (nuclear matrix and cytoskeletal elements) from cultured cells. This method's main feature is that cell fractionation is performed in situ. Structures not solubilized in a particular extraction step remain attached to the substrate and retain their morphology. Before and after each extraction step they can be analyzed for the presence and location of the protein under study by using immunological or cytochemical techniques. In this way the in vivo origin of a protein solubilized in a particular extraction step is determined. The solubilized protein then may be further characterized biochemically. To allow analyses of proteins associated with the residual cellular skeleton, conditions were developed for its solubilization that do not interfere with enzymatic and immunological studies.

Accession: 006173593

PMID: 6725403

DOI: 10.2307/1610714

Download PDF Full Text: Preparation of nuclear matrices from cultured cells subfractionation of nuclei in situ



Related references

Localization of nuclear antigens during preparation of nuclear matrices in situ. Canadian Journal of Biochemistry and Cell Biology 63(6): 644-653, 1985

Spatial organization of four hnRNP proteins in relation to sites of transcription, to nuclear speckles, and to each other in interphase nuclei and nuclear matrices of HeLa cells. Experimental Cell Research 246(2): 461-470, 1999

Spatial organization of four hnRBP proteins in relation to sites of transcription, to nuclear speckles, and to each other in interphase nuclei and nuclear matrices of HeLa cells. Experimental Cell Research 246(2): 461-470, Feb 1, 1999

Isolation and fractionation of nuclear matrices from cultured cells. Tsitologiya 28(8): 843-847, 1986

Preparation of extracellular matrices produced by cultured bovine corneal endothelial cells and pf hr 9 endodermal cells their use in cell culture. Barnes, D W , D A Sirbasku And G H Sato (Ed ) Cell Culture Methods For Molecular And Cell Biology, Vol 1 Methods For Preparation Of Media, Supplements, And Substrata For Serum-Free Animal Cell Culture Xxiv+353p Alan R Liss, Inc : New York, N Y , Usa Illus 275-294, 1984

Studies on plastid nuclei nucleoids in nicotiana tabacum l. i. isolation of proplastid nuclei from cultured cells and identification of proplastid nuclear proteins. Plant and Cell Physiology 29(1): 167-178, 1988

Preparation of extracellular matrices produced by cultured corneal endothelial and PF-HR9 endodermal cells. Current Protocols in Cell Biology Chapter 10: Unit 10.4-Unit 10.4, 2008

Nuclear cortex dissociation of nuclei isolated from cultured human cells. Biologie Cellulaire (Ivry Sur Seine) 36(1): 87-90, 1979

Isolation of sperm nuclei and nuclear matrices from the mouse, and other rodents. Methods in Molecular Biology 927: 437-444, 2013

Simultaneous isolation of cell-nuclei, plastid-nuclei and mitochondrial-nuclei from cultured tobacco cells; comparative analysis of their transcriptional activities in vitro. Plant Science (Shannon) 133(1): 17-31, April 6, 1998