PMID: 4287141Nov 1, 1965Paper

Isolation of skeletal muscle nuclei

The Journal of Cell Biology
J C EdelmanI L Schwartz

Abstract

A method employing aqueous media for isolation of nuclei from rat skeletal muscle is described. The technique involves (a) mincing and then homogenizing in a 0.32 M sucrose-salt solution with a Potter-Elvehjem type homogenizer using a Delrin (an acetal resin) pestle and a carefully controlled, relatively large pestle-to-glass clearance, (b) filtering through fiberglass and stainless steel screens of predetermined mesh size to remove myofibrils and connective tissue, and (c) centrifuging in a 2.15 M sucrose-salt solution containing 0.7 mM ATP. Electron and phase-contrast microscopic observations show that the nuclei are intact, unencumbered by cytoplasmic tags, and possess well preserved distinct nucleoli, nucleoplasm, and nuclear membranes. Cytoplasmic contamination is minimal and mainly mitochondrial. Chemical assays of the nuclear fraction show that the DNA/protein and RNA/DNA ratios are comparable to those obtained in other tissues. These ratios, as well as the low specific activity obtained for cytochrome c oxidase and the virtual absence of myofibrillar ATPase, indicate a high degree of purity with minimal mitochondrial and myofibrillar contamination. The steps comprising the technique and the reasons for their selection a...Continue Reading

References

Jan 20, 1957·The Journal of General Physiology·V G ALLFREYS OSAWA
Oct 1, 1962·The Journal of Cell Biology·M B SPORNW DINGMAN
May 1, 1964·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·W C HYMER, E L KUFF
Jan 1, 1952·The Journal of General Physiology·V ALLFREYH SAETREN
Jan 1, 1952·The Journal of General Physiology·H STERNH SAETREN

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Citations

Jul 1, 1979·Journal of the Neurological Sciences·I H FraserC C Liew
Mar 1, 1967·The Journal of Cell Biology·C C WidnellJ R Tata
Apr 1, 1968·The Journal of Cell Biology·P D Sadowski, J W Steiner
Jul 1, 1969·The Journal of Cell Biology·W W Franke, W Schinko
Mar 15, 1975·Experimental Cell Research·L Kuehl
Apr 1, 1970·Archives of Biochemistry and Biophysics·B E Sobel, S Kaufman
May 1, 1966·The American Journal of Medicine·P M Edelman, I L Schwartz
Apr 5, 2011·Cell Metabolism·Péter BaiJohan Auwerx
Dec 12, 2013·American Journal of Physiology. Endocrinology and Metabolism·Veeraj GoyaramEdward O Ojuka
May 24, 2006·Human Molecular Genetics·Xiaoyan LinCharles A Thornton

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