| Abbreviations | p. vii |
| Preface | p. ix |
| Principles and strategies of centrifugation | p. 1 |
| Sedimentation of particles under the influence of gravity | p. 1 |
| Rate of sedimentation | p. 2 |
| Increasing the magnitude of the gravitational field | p. 2 |
| Differential centrifugation | p. 3 |
| Centrifugation in density gradients | p. 5 |
| Sedimentation coefficients and the Svedberg equation | p. 12 |
| Centrifugation hardware | p. 15 |
| Centrifuges--an overview | p. 15 |
| Basic rotor design | p. 17 |
| Rotor capacity and performance | p. 18 |
| Uses of centrifuges and rotors | p. 19 |
| Centrifuge rotor parameters | p. 19 |
| Centrifuge tubes | p. 21 |
| Good tube and rotor practice | p. 25 |
| Sedimentation of particles in rotors | p. 28 |
| Rotors for preformed density gradients | p. 33 |
| Zonal rotors | p. 35 |
| The analytical ultracentrifuge | p. 39 |
| Gradient media | p. 43 |
| Choosing a suitable density-gradient medium | p. 43 |
| Physicochemical characteristics of gradient media | p. 47 |
| Preparing solutions of gradient media | p. 51 |
| Analysis of fractions | p. 56 |
| Gradient techniques | p. 61 |
| Types of gradients and their uses | p. 61 |
| Pre-formed discontinuous gradients | p. 62 |
| Pre-formed continuous gradients | p. 64 |
| Nonlinear gradients | p. 69 |
| Self-generating gradients | p. 72 |
| Harvesting gradients | p. 76 |
| Purification of mammalian cells | p. 85 |
| Introduction | p. 85 |
| Selection of medium | p. 85 |
| Handling of gradient media | p. 86 |
| Centrifugation strategy | p. 88 |
| Peripheral blood mononuclear cells | p. 90 |
| Human monocytes | p. 91 |
| Dendritic cells | p. 93 |
| Isolation of Islets of Langerhans from porcine pancreas | p. 93 |
| Stellate cells from mammalian liver | p. 94 |
| Other low-density cells | p. 95 |
| Purification of mononuclear cells by flotation from whole blood (human, rat and mouse) | p. 97 |
| Purification of monocytes by flotation from whole human blood | p. 98 |
| Purification of dendritic cells by flotation from mouse tissues | p. 99 |
| Purification of Islets of Langerhans by flotation from digested porcine pancreas | p. 100 |
| Purification of stellate cells from mammalian liver | p. 101 |
| Fractionation of subcellular organelles | p. 103 |
| Introduction | p. 103 |
| Homogenization of tissue or cells | p. 103 |
| Physical characteristics of subcellular particles | p. 108 |
| Differential centrifugation of a rat liver homogenate | p. 110 |
| Purification of organelles in gradients | p. 114 |
| Isolation of rat liver mitochondria by differential centrifugation | p. 123 |
| Isolation of rat liver nuclei in a sucrose gradient | p. 125 |
| Isolation of rat liver nuclei in a iodixanol gradient | p. 127 |
| Isolation of rat liver mitochondria in a Percoll gradient | p. 128 |
| Isolation of rat liver peroxisomes in a pre-formed iodixanol gradient | p. 130 |
| Isolation of rat liver lysosomes in a Percoll gradient | p. 132 |
| Isolation of rat liver lysosomes in a Nycodenz gradient | p. 133 |
| Rapid isolation of 'stacked' rat liver Golgi on a sucrose barrier | p. 134 |
| Fractionation of a light mitochondrial fraction in a self-generated iodixanol gradient | p. 136 |
| Isolation of plasma membrane sheets from rat liver | p. 138 |
| Fractionation of membrane vesicles | p. 141 |
| The membrane compartments | p. 141 |
| Density and size of membrane vesicles | p. 142 |
| Separation strategies | p. 143 |
| Endoplasmic reticulum | p. 145 |
| Golgi membranes | p. 146 |
| Plasma membrane and membrane domains | p. 146 |
| Isolation of vesicular membranes by density perturbation | p. 148 |
| Analytical gradients (secretion) | p. 151 |
| Analytical gradients (endocytosis) | p. 153 |
| Density barrier separation of RER and SER | p. 159 |
| Separation of SER and RER on a continuous sucrose gradient | p. 160 |
| Isolation of Golgi membrane vesicles from an homogenate in a discontinuous sucrose gradient | p. 162 |
| Isolation of the hepatocyte sinusoidal membrane domain using immunoaffinity beads | p. 163 |
| Analysis of the secretory process in cultured cells using discontinuous sucrose-D[subscript 2]O density gradients | p. 165 |
| Sedimentation velocity gradients | p. 166 |
| Fractionation of Golgi, SER and RER in a self-generated iodixanol gradient | p. 167 |
| Fractionation of macromolecules and macromolecular complexes | p. 169 |
| Buoyant density banding of nucleic acids and proteins | p. 169 |
| Rate-zonal (sedimentation-velocity) banding of proteins and ribonucleoproteins | p. 173 |
| Viruses | p. 178 |
| Plasma lipoproteins | p. 183 |
| Purification of plasmid DNA from bacteria using CsCl-EtBr | p. 188 |
| Purification of plasmid DNA using DAPI-iodixanol gradients | p. 190 |
| Sedimentation velocity analysis of proteins in a 5-ml pre-formed sucrose gradient | p. 191 |
| Small-scale sedimentation velocity analysis of proteins in a self-generated iodixanol gradient | p. 192 |
| Isolation of polysomes from E. coli | p. 194 |
| Isolation of rat liver ribosomal subunits | p. 196 |
| Concentration and purification of Sendai virus (100-200 ml) using a large volume fixed-angle rotor | p. 198 |
| Concentration and purification of rAAV in a discontinuous iodixanol gradient | p. 199 |
| Concentration and purification of virus using a self-generated iodixanol gradient | p. 200 |
| Fractionation of lipoproteins by sequential flotation using KBr | p. 201 |
| Fractionation of lipoproteins in self-generated gradients of iodixanol | p. 203 |
| List of manufacturers and technical support sources | p. 205 |
| Index | p. 207 |
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