Isolation of a Golgi Apparatus-Rich Fraction From Rat Liver

Morré, D. James; Hamilton, Robert L.; Mollenhauer, Hilton H.; Mahley, Robert W.; Cunningham, William P.; Cheetham, R. D.; LeQuire, Virgil S.

doi:10.1083/jcb.44.3.484

Cited by 151 publications

(30 citation statements)

References 11 publications

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“…A Golgi-rich fraction was obtained from rat liver following a modification of the method of Morrd et al (28) introduced by Wibo et al? Peroxisomes were prepared as described by Leighton et al (24).…”

Section: Preparation Of Subcellular Membrane Fractionsmentioning

confidence: 99%

Analytical study of microsomes and isolated subcellular membranes from rat liver. V. Immunological localization of cytochrome b5 by electron microscopy: methodology and application to various subcellular fractions.

Fowler¹,

Remacle²,

Trouet³

et al. 1976

The Journal of Cell Biology

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The localization of cytochrome b5 on the membranes of various subcellular organelles of rat liver was studied by a cytoimmunological procedure using anticytochrome bdanti-ferritin hybrid antibodies and ferritin as label. For this study, highly purified and biochemically characterized membrane preparations were employed. Outer mitochondrial membranes were found to be heavily labeled by the hybrid antibodies whereas Golgi and plasma membranes were not marked by the reagent. Peroxisome membranes were moderately labeled by the hybrid antibodies, suggesting that they may contain some cytochrome bs. The preparation and purification of hybrid antibodies without peptic digestion is described and an analysis made of the composition of the final reagent product.Cytochrome b5 was first found in the microsomal fraction of rat liver (10, 45) and later shown to be associated with outer membranes of mitochondria (17,33,43). This protein has also been reported to be present in the Golgi membranes (8,16,21) and NADH: cytochrome b5 reductase activity is said to be present in peroxisomes (9, 13). On the basis of these data, cytochrome bs would appear to be present in membranes of several of the organelles of the liver cell. However, in these biochemical studies it is not always certain that the cytochrome b~ detected is truly associated with the organelle under study rather than being merely present as a subcellular contaminant of the preparations analyzed. In this paper, we report a sensitive ferritin-hybrid antibody technique that does permit, by morphological analysis, the specific localization of cytochrome b5 to individual membrane elements. In the course of developing this assay, we studied in detail the various steps in the preparation of our morphologic label, an antigenantibody complex between ferritin and the anticytochrome bdanti-ferritin hybrid antibody (abd aF). 1 We describe here our experience concerning 1 Abbreviations used in this paper: abdaF-ferritin, antigen-antibody complex between ferritin and the anti-cytochrome bs/anti-ferritin hybrid antibody; anti-X, an immunoglobulin or a fragment of an immunoglobulin, the specificity of which is undefined; p, density; ER, endoplasmic reticulum; PBS, 0.01 M Na phosphate, pH 7.4, in 0.15 M NaCI; SDS, sodium dodecyl sulfate.

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Section: Preparation Of Subcellular Membrane Fractionsmentioning

confidence: 99%

Analytical study of microsomes and isolated subcellular membranes from rat liver. V. Immunological localization of cytochrome b5 by electron microscopy: methodology and application to various subcellular fractions.

Fowler¹,

Remacle²,

Trouet³

et al. 1976

The Journal of Cell Biology

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show abstract

“…The Golgi apparatus is an organelle which appears and, more recently, of negatively stained preparato contribute both to the development and to the tions from isolated Golgi fractions (3) .I From normal function of the liver cell . The complex I Sturgess, J .…”

Section: Introductionmentioning

confidence: 99%

Morphometry of the Golgi Apparatus in Developing Liver

Sturgess

Iglesia

1972

The Journal of Cell Biology

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“…3) showed a system of tubules and plate -like center regions as decribed in the Golgi fraction from rat liver (Fleischer et al . 1969;Morre et al 1970). …”

Section: Discussionmentioning

confidence: 99%