Document Detail

Large-scale isolation of Cajal bodies from HeLa cells.
MedLine Citation:
PMID:  12134083     Owner:  NLM     Status:  MEDLINE    
The Cajal body (CB) is a conserved, dynamic nuclear structure that is implicated in various cellular processes, such as the maturation of splicing small nuclear ribonucleoproteins and the assembly of transcription complexes. Here, we report the first procedure for the large-scale purification of CBs from HeLa cell nuclei, resulting in an approximately 750-fold enrichment of the CB marker protein p80-coilin. Immunofluorescence, immunoblotting, and mass spectrometric analyses showed that the composition of the isolated CBs was similar to that of CBs in situ. The morphology and structure of the isolated CBs, as judged by transmission and scanning electron microscopy analysis, are also similar to those of CBs in situ. This protocol demonstrates the feasibility of isolating intact distinct classes of subnuclear bodies from cultured cells in sufficient yield and purity to allow detailed characterization of their molecular composition, structure, and properties.
Yun Wah Lam; Carol E Lyon; Angus I Lamond
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular biology of the cell     Volume:  13     ISSN:  1059-1524     ISO Abbreviation:  Mol. Biol. Cell     Publication Date:  2002 Jul 
Date Detail:
Created Date:  2002-07-22     Completed Date:  2003-05-07     Revised Date:  2013-04-18    
Medline Journal Info:
Nlm Unique ID:  9201390     Medline TA:  Mol Biol Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2461-73     Citation Subset:  IM    
Wellcome Trust Biocentre, MSI/WTB Complex, University of Dundee, Dundee DD1 4HN, Scotland, United Kingdom.
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MeSH Terms
Cell Fractionation
Cell Nucleus / chemistry,  metabolism*
Coiled Bodies / chemistry*,  metabolism,  ultrastructure
HeLa Cells
Magnesium / metabolism
Nuclear Proteins / isolation & purification*,  metabolism
Reg. No./Substance:
0/Nuclear Proteins; 136882-81-0/p80-coilin; 7439-95-4/Magnesium

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