Fractionation of Membrane‐Bound Polysomes, Free Polysomes, and Nuclei from Tissue‐Cultured Cells

Abstract

Free polysomes from many kinds of eukaryotic cells are liberated by cell lysis using the non‐ionic detergent Kyro EOB leaving a residue of nuclei with strands of rough endoplasmic reticulum attached. Subsequent treatment with a combination of Kyro EOB plus deoxycholate solubilizes the membrane‐bound polysomes. Polysomes prepared in this manner function well in amino‐acid‐incorporating systems in vitro. The nuclei remaining incorporate [ 14 C]ATP into RNA. More active nuclei, however, are obtained when cells are disrupted by nitrogen cavitation.