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Several chemical methods for the isolation of cell nuclei have recently been described. Calf thymus nuclei have been isolated by mild hypotonic shock followed by Ficoll gradient purification (1). A nonaqueous technique has been employed by Hammel and Bessman for purifying duck erythrocyte nuclei (2). Cell nuclei from cat brain have been obtained by using a detergent, Tri ton X-100, which results in a 2.8-fold enrichment of nuclei (as measured by the increase in the ratio of DNA/prote in) (3). Tri ton X-100 has also been used to isolate rat liver nuclei, and in this preparation about 5-fold enrichment was obtained (4). Citric acid at pH 3.5 has been used to prepare HeLa cell nuclei, which were reported to appear uncontaminated by cytoplasmic material under the phase microscope (5). In the present report the isolation of HeLa cell nuclei with sodium deoxycholate is described. The purity of the nuclei was evaluated by comparing the DNA-to-protein ratios of the whole cells to those of the separated cell fractions (6). The activity of fl-glucuronidase was used as a measure of the lysosomal contamination of the various cell fractions. Assays of these cell fractions revealed that most of the alkaline phosphatase activity was associated with the nuclear fraction. M A T E R I A L S A N D M E T H O D S

the journal of cell biology/the journal of cell biology

A CHEMICAL METHOD FOR THE ISOLATION OF H<scp>E</scp>L<scp>A</scp> CELL NUCLEI AND THE NUCLEAR LOCALIZATION OF H<scp>E</scp>L<scp>A</scp> CELL ALKALINE PHOSPHATASE

A CHEMICAL METHOD FOR THE ISOLATION OF HELA CELL NUCLEI AND THE NUCLEAR LOCALIZATION OF HELA CELL ALKALINE PHOSPHATASE