Isolation of ribosomes and polysomes.
Here we describe a preparative differential centrifugation protocol for the isolation of ribosomes from a crude cell homogenate. The subcellular fraction obtained is enriched in ribosome monomers and polysomes. The protocol has been optimized for the homogenization and collection of the ribosomal fraction from prokaryotic cells, mammalian and plant tissues, reticulocytes, and chloroplasts. The quality of the ribosomal preparation is enhanced by the removal of the remaining cellular components and adsorbed proteins by pelleting through a sucrose cushion with a high concentration of monovalent salts, NH4Cl or KCl. The different components of the ribosomal fraction isolated using this protocol can be further purified by sucrose gradient centrifugation.
Department of Biology and Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284;
Maria C
MC
Rivera
Primary Pharmacology Group, Pfizer Global Research and Development, Groton, Connecticut 06340;
Bruce
B
Maguire
Department of Molecular, Cell and Developmental Biology and Department of Human Genetics, University of California, Los Angeles, California 90095.
James A
JA
Lake
D002498:Centrifugation / Q000379:methods*; D045424:Complex Mixtures; D005057:Eukaryotic Cells / Q000737:chemistry; D011387:Prokaryotic Cells / Q000737:chemistry; D012270:Ribosomes
D045424:Complex Mixtures