DETERGENT LYSIS FOR ISOLATION OF INTACT POLYSOMES OF NOVIKOFF HEPATOMA ASCITES-CELLS

  • 1 January 1976
    • journal article
    • research article
    • Vol. 36  (3) , 1026-1034
Abstract
An efficient, rapid procedure for the isolation of polysomes from [rat] Novikoff hepatoma ascites cells that avoids mechanical shear was developed, and Ivory detergent was used to lyse the cells. This method yields classes of large polysomes and 3 to 5-fold increased yields over that of routine homogenization procedures. Up to 70% of the polysome population is larger than tetramers. The polysomes and ribosomes are morphologically intact and contain undegraded rRNA and biologically active mRNA. This procedure also yields nuclei free of cytoplasmic contamination as a by-product of polysome isolation. These nuclei do not show evidence of protein degradation and are enzymatically active with respect to RNA polymerases I and II.

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