This article describes a method for the isolation of nuclei from guinea pig liver. It involves the homogenization of the tissue in 0.88 M sucrose-1.5 mM CaCl2 followed by centrifugation in a discontinuous density gradient in which the upper phase is the homogenate and the lower phase is 2.2 M sucrose-0.5 mM CaCl2. Based on DNA recovery, the isolated fraction contains 25 to 30 per cent of the nuclei of the original homogenate. Electron microscopical observations showed that ∼88 per cent of the isolated nuclei come from liver cells (the rest from von Kupffer cells and leucocytes) and that ∼90 per cent of the nuclei appear intact, with well preserved nucleoli, nucleoplasm, nuclear envelope, and pores. Cytoplasmic contamination is minimal and consists primarily of the nuclear envelope and its attached ribosomes. The nuclear fraction consists of ∼22.3 per cent DNA, ∼4.7 per cent RNA, and ∼73 per cent protein, the DNA/RNA ratio being 4.7. Data on RNA extractibility by phosphate and salt and on the base composition of total nuclear RNA are included.
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August 01 1963
STUDIES ON ISOLATED NUCLEI : I. Isolation and Chemical Characterization of a Nuclear Fraction from Guinea Pig Liver
Rachele Maggio,
Rachele Maggio
From The Rockefeller Institute
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Philip Siekevitz,
Philip Siekevitz
From The Rockefeller Institute
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George E. Palade
George E. Palade
From The Rockefeller Institute
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Rachele Maggio
From The Rockefeller Institute
Philip Siekevitz
From The Rockefeller Institute
George E. Palade
From The Rockefeller Institute
Received:
January 10 1963
Online ISSN: 1540-8140
Print ISSN: 0021-9525
Copyright © 1963 by The Rockefeller Institute Press
1963
J Cell Biol (1963) 18 (2): 267–291.
Article history
Received:
January 10 1963
Citation
Rachele Maggio, Philip Siekevitz, George E. Palade; STUDIES ON ISOLATED NUCLEI : I. Isolation and Chemical Characterization of a Nuclear Fraction from Guinea Pig Liver . J Cell Biol 1 August 1963; 18 (2): 267–291. doi: https://doi.org/10.1083/jcb.18.2.267
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