A comparison of methods for the isolation and fractionation of reticulocyte ribosomes

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Abstract
Polysomes, ribosomes and pH 5 enzymes were isolated from rabbit reticulocytes by acidifying the post-mitochondrial supernatant to pH 6.0 to precipitate all ribonucleo-protein particles and about half the pH 5 enzymes; the precipitate was redissolved in buffer, pH 7.6, and fractionated by zone centrifuging. The isolation of polysome-rich and ribosome-rich fractions from the post-mitochondrial supernatant was also examined. Studies of the stability of polysomes revealed that dissociation into sub-units occurred when both bound and free Mg2+ was chelated by EDTA or when the pH was increased above pH 8.8.