Batchwise Purification of Liver Ribosomes and Polysomes from Unfasted Mice on Hydroxylapatite |
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Authors: | W. L. Hoffman J. Ilan |
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Affiliation: | Department of Anatomy , School of Medicine Case Western Reserve University , Cleveland, Ohio, 44106 |
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Abstract: | Batchwise purification of liver ribosomes and polysomes on hydroxyl-apatite is a rapid procedure to remove glycogen, hemoglobin, ribonuclease and other contaminants from ribosomal preparations. Ribosomes and polysomes are adsorbed to hydroxylapatite in a Büchner filter funnel and the contaminants are eluted from the hydroxylapatite with 0.15 M KH2PO4. The ribosomes and polysomes are then eluted with 0.3–0.4 M KH2PO4 and concentrated by centrifugation. The resolution of the polysome profiles was greatly improved following purification. The purified ribosomes could be dissociated into subunits at 0.3 M KCl, and showed no loss of activity in poly-U directed phenylalanine synthesis. |
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