[Cancer Research 30, 352-356, February 1, 1970]
© 1970 American Association for Cancer Research
A Rapid Method for the Isolation of Nuclei from Ehrlich Ascites Tumor Cells1
F. P. Mamaril,
Areta Dobrjansky and
Saul Green
Division of Biochemistry, Sloan-Kettering Institute for Cancer Research, New York, New York 10021
A method for the isolation of nuclei from Ehrlich ascites tumor cells free of cytoplasmic contamination is described. The procedure involves hypotonic osmotic shock, homogenization with a close-fitting Dounce homogenizer, and differential centrifugation through two different high-density sucrose solutions. The extent of cytoplasmic contamination of the isolated nuclei at each stage was followed by DNA, RNA, protein, and marker enzyme analysis. The final product was examined by phase and electron microscopy and indicated the presence of a high yield of morphologically intact Ehrlich ascites tumor cell nuclei, free of cytoplasmic tags.
Isolated ascites cell nuclei contained nicotinamide adenine dinucleotide glycohydrolase (EC 3.2.2.5) which could not be solubilized by lipase treatment and which differed significantly from the microsomal enzyme in its pH optimum and heat stability.
1 This work was supported in part by Grant CA-08748 from the National Cancer Institute, NIH, and Grant T-431K from the American Cancer Society.
Received 4/14/69.
Accepted 7/17/69.
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cDNA-derived molecular characteristics and antibodies to a new centrosome-associated and G2/M phase-prevalent protein
J. Cell Sci.,
January 1, 1993;
104(1):
19 - 30.
[Abstract]
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Copyright © 1970 by the American Association for Cancer Research.
