Zonal rotor ultracentrifugation: a proposal for improving the effectiveness of the procedure

1. Purified liver nuclei from adult rats separate into two main zones when centrifuged in the slow-speed zonal rotor. One zone contains diploid nuclei, the other tetraploid. 2. The effect of age on the pattern of rat liver ploidy was examined. Tetraploid nuclei are virtually absent from young animals. They increase in proportion steadily with age. Partial hepatectomy disturbs the pattern of ploidy. 3. The zonal centrifuge permits the separation of diploid, tetraploid, octaploid and hexadecaploid nuclei from mouse liver. 4. Rat liver nuclei are isopycnic with sucrose solutions of density 1·35 at 5°.

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Large-scale preparation of synaptosomes from bovine brain using a zonal rotor technique

Neurochemical Research ◽

10.1007/bf00964643 ◽

1979 ◽

Vol 4 (4) ◽

pp. 483-504 ◽

Cited By ~ 15

Author(s):

Kenneth C. Leskawa ◽

Herbert C. Yohe ◽

Michiko Matsumoto ◽

Abraham Rosenberg

Keyword(s):

Large Scale ◽

Bovine Brain ◽

Large Scale Preparation ◽

Scale Preparation ◽

Zonal Rotor

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Large Capacity Separation of Cells in a Reorienting Zonal Rotor

Cell Separation ◽

10.1016/b978-0-12-564502-7.50018-9 ◽

1983 ◽

pp. 221-233 ◽

Cited By ~ 1

Author(s):

THERESA P. PRETLOW ◽

THOMAS G. PRETLOW

Keyword(s):

Large Capacity ◽

Zonal Rotor

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Enzyme distribution in mammalian membranes**The following abbreviations are used: ER, endoplasmic reticulum (excluding the Golgi); PM, plasma membrane (the equivalent term ‘cell membrane’ is not favoured). The usage of the terms ‘cytochemical’ (here restricted to EM studies), ‘microsome’ and ‘microsomal [or mitochondrial, lysosomal, etc.] fraction’ is indicated in the text. Membranes in apposition are best termed ‘paired', not ‘double’. A ‘vesicle’ is a ‘small bladder-like vessel’ or ‘sac’ (Oxford Dictionary). The centrifugal procedure commonly termed ‘differential centrifugation’ is here termed ‘differential pelleting’, in contrast with ‘banding’ or ‘zonal centrifugation’; the latter term does not necessarily imply use of a zonal rotor (Reid, 1972a; Reid and Williamson, 1974).

Mammalian Cell Membranes ◽

10.1016/b978-0-408-70722-0.50011-5 ◽

1976 ◽

pp. 161-197 ◽

Cited By ~ 2

Author(s):

R.H. Hinton ◽

E. Reid

Keyword(s):

Endoplasmic Reticulum ◽

Plasma Membrane ◽

Cell Membrane ◽

Differential Centrifugation ◽

Enzyme Distribution ◽

Oxford Dictionary ◽

Zonal Rotor

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A ZONAL ROTOR METHOD FOR THE PREPARATION OF MICROPEROXISOMES FROM EPITHELIAL CELLS OF GUINEA PIG SMALL INTESTINE

The Journal of Cell Biology ◽

10.1083/jcb.61.1.123 ◽

1974 ◽

Vol 61 (1) ◽

pp. 123-133 ◽

Cited By ~ 24

Author(s):

M. J. Connock ◽

P. R. Kirk ◽

A. P. Sturdee

Keyword(s):

Small Intestine ◽

Electron Microscope ◽

Epithelial Cells ◽

Guinea Pig ◽

Analytical Data ◽

Microscope Examination ◽

Electron Microscope Examination ◽

Large Particles ◽

Zonal Separation ◽

Zonal Rotor

A method is described for the preparation of catalase particles from homogenates made from suspensions of epithelial cells of the small intestine of the guinea pig. Electron microscope examination of the preparations revealed the presence of small diaminobenzidine-positive particles measuring 0.1–0.3 nm in diameter and resembling the microperoxisomes observed by Novikoff and Novikoff (1972. J. Cell Biol. 53:532.). Analytical data upon which the method is based are presented. The technique consisted of a rate zonal separation of microperoxisomes from large particles followed by an isopycnic separation from less dense organelles. Application of the method yielded microperoxisomes purified between 20- and 30-fold.

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A study of nucleated erythrocytes by density-gradient centrifugation in a zonal rotor

Biochemical Journal ◽

10.1042/bj1110583 ◽

1969 ◽

Vol 111 (4) ◽

pp. 583-591 ◽

Cited By ~ 11

Author(s):

A P Mathias ◽

D. Ridge ◽

N. St G. Trezona

Keyword(s):

Molecular Weight ◽

Ribosomal Rna ◽

Base Composition ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Density Gradients ◽

Avian Erythrocytes ◽

Nuclear Rna ◽

Zonal Rotor

1. Several substances of high molecular weight were examined for their suitability as suspension media in the formation of density gradients for the zonal centrifugation of avian erythrocytes. None proved satisfactory. 2. The behaviour of pigeon erythrocytes in rate-sedimentation experiments in a type A zonal rotor with density gradients of sucrose was examined. The mature cells sediment more rapidly than the younger cells and have a lower RNA/DNA ratio. Maturation is accompanied by a greater loss of RNA from the nucleus than from the cytoplasm. 3. The base composition of the nuclear RNA and of the two species of cytoplasmic ribosomal RNA is reported. 4. The RNA of erythrocytes may be labelled in vivo by injection of inorganic [32P]phosphate. The cells most active in the synthesis of RNA sediment less rapidly than the bulk of the cells. 5. Reticulocyte nuclei sediment more slowly than those from erythrocytes. Reticulocyte nuclei have a mean volume of 35μ3 and are isopycnic with sucrose of density 1·2871 (measured at 20°). Maturation of the nuclei causes them to shrink to a volume of 25μ3 and the density to increase to 1·2944.

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