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Isolation and characterization of nuclear lamina from ehrlich ascites tumor cells

✍ Scribed by Chavdar Krachmarov; Bistra Tasheva; Dimitar Markov; Ronald Hancock; George Dessev

Publisher
John Wiley and Sons
Year
1986
Tongue
English
Weight
923 KB
Volume
30
Category
Article
ISSN
0730-2312

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✦ Synopsis

We have developed a simple and rapid method for isolation of purified nuclear lamina from Ehrlich ascites tumor cells. The procedure employs chromatin structures prepared from whole cells at low ionic strength and is carried out under conditions that minimize the formation of artifactual protein-DNA complexes. When the isolation is performed in the presence of EDTA, nuclear lamina without distinct pore complexes is obtained. In the absence of EDTA, intact pore complexes and a large amount of vimentin 100 A filaments are seen associated with nuclear lamina. The main nuclear lamina proteins are characterized using gel electrophoresis, immunoblotting, and two-dimensional peptide mapping. An extensive structural homology is found between lamin A and lamin C, whose peptide maps differ by only one major spot, whereas lamin B has apparently unrelated pattern.

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