scholarly journals ISOLATION AND FRACTIONATION OF RAT BRAIN NUCLEI

1966 ◽  
Vol 30 (2) ◽  
pp. 405-415 ◽  
Author(s):  
H. Løvtrup-Rein ◽  
B. S. McEwen
Keyword(s):  
Rat Brain ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Differential Centrifugation ◽  
Brain Nuclei ◽  
Isolated Nuclei ◽  
Isolation Media

A method for isolating pure and unaltered nuclei from rat brain by means of differential centrifugation is described. The isolated nuclei are further separated into discrete fractions of neuronal, astrocytic, and glial nuclei, with a yield amounting to 20 to 25% of the DNA of the original homogenate. Both the morphology and size of the nuclei remained unchanged. Problems concerning the composition of the isolation media, the use of detergents, as well as those raised by density gradient centrifugation in sucrose, Ficoll, and Dextran are discussed. Some values for the density of each type of brain nuclei are suggested.

scholarly journals Intramitochondrial localization of δ-aminolaevulate synthetase and ferrochelatase in rat liver

1969 ◽  
Vol 114 (3) ◽  
pp. 455-461 ◽  
Author(s):  
Roxane McKay ◽  
R. Druyan ◽  
G. S. Getz ◽  
M. Rabinowitz
Keyword(s):  
Glutamate Dehydrogenase ◽  
Density Gradient ◽  
Malate Dehydrogenase ◽  
Rat Liver ◽  
Mitochondrial Biogenesis ◽  
Mitochondrial Membrane ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Differential Centrifugation ◽  
Inner Mitochondrial Membrane

Intramitochondrial loci for δ-aminolaevulate synthetase and ferrochelatase, the initial and final enzymes in haem synthesis, have been found in rat liver. Two different methods of fractionation were applied to mitochondria: (a) sonication and density-gradient centrifugation; (b) treatment with digitonin and differential centrifugation. Similar results were obtained with each technique. δ-Aminolaevulate synthetase is distributed similarly to two known matrix enzymes, malate dehydrogenase and glutamate dehydrogenase. Ferrochelatase is firmly bound to the the inner mitochondrial membrane. These results are considered in terms of the regulation of haem synthesis and in relation to mitochondrial biogenesis.

scholarly journals The locations of cathepsin activity and β-glucuronidase in the Guerin T8 tumour

1970 ◽  
Vol 118 (3) ◽  
pp. 543-549 ◽  
Author(s):  
A. R. Poole
Keyword(s):  
Endoplasmic Reticulum ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Differential Centrifugation ◽  
Unit Membrane ◽  
Electron Microscopic ◽  
Cathepsin Activity ◽  
Density Gradient Sedimentation

Tumour homogenate fractions, isolated by differential centrifugation, were subfractionated by density-gradient centrifugation. Biochemical and electron microscopic analyses revealed that β-glucuronidase and cathepsin activity were associated with a class (possibly two) of lysosomal particles of density greater than those of mitochondria and the endoplasmic reticulum. Lysosomes sedimented by low g forces were vacuolar, electron-dense, delineated by a unit membrane and about 0.2μm in diameter. β-Glucuronidase was also apparently associated with ribosomes whereas cathepsin was bound in part to the endoplasmic reticulum. Catalase and glucose 6-phosphatase possessed slightly different density-gradient sedimentation profiles.

scholarly journals Some Properties of Rose Mosaic Virus from South Australia

1970 ◽  
Vol 23 (5) ◽  
pp. 1197 ◽  
Author(s):  
AA Basit ◽  
RIB Francki
Keyword(s):  
North America ◽  
Physical Properties ◽  
Mosaic Virus ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
South Australia ◽  
Sucrose Density Gradient ◽  
Differential Centrifugation ◽  
Sucrose Density Gradient Centrifugation

Isolates of rose mosaic virus (RMV) from South Australia were purified by differential centrifugation of cucumber extracts clarified by emulsification with ether, followed by sucrose density-gradient centrifugation. The virus was shown to be serologically similar to and to have many physical properties in common with RMV from North America. However, the Australian isolates studied appear to hlwe narrower host ranges.

Isolation of the 67Gallium Accumulating Fraction in Normal Rat Liver

1974 ◽  
Vol 13 (01) ◽  
pp. 72-84
Author(s):  
K. Hierholzer ◽  
K. zum Winkel ◽  
U. Haubold ◽  
E. Aulbert
Keyword(s):  
Electron Microscopy ◽  
Density Gradient ◽  
Intravenous Injection ◽  
Rat Liver ◽  
Soluble Fraction ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Differential Centrifugation ◽  
Normal Rat

SummarySubcellular 67Gallium distribution was investigated in normal rat liver after intravenous injection. By differential centrifugation and density gradient centrifugation 67Gallium accumulating bodies were isolated and identified as lysosomes by enzyme determination and electron microscopy. 67Gallium enrichment in this fraction was 23-fold. Using the isolated 67Gallium accumulating lysosomes the binding state of the isotope inside the lysosomes was studied. 67Gallium was found to be associated with the soluble fraction of lysosomes.

Identification of Mineralocorticoid Binding Sites in Rat Brain by Competition Studies and Density Gradient Centrifugation

1983 ◽  
Vol 37 (5) ◽  
pp. 354-360 ◽  
Author(s):  
Hector Coirini ◽  
Elisa T. Marusic ◽  
Alejandro F. De Nicola ◽  
Thomas C. Rainbow ◽  
Bruce S. McEwen
Keyword(s):  
Rat Brain ◽  
Density Gradient ◽  
Binding Sites ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation

scholarly journals Collagenolytic cathepsin activity in rabbit peritoneal polymorphonuclear leucocyte granules

1978 ◽  
Vol 172 (1) ◽  
pp. 83-89 ◽  
Author(s):  
W T Gibson ◽  
D W Milsom ◽  
F S Steven ◽  
J S Lowe
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Polymorphonuclear Leucocyte ◽  
Polymorphonuclear Leucocytes ◽  
Differential Centrifugation ◽  
Ph Optimum ◽  
Cathepsin Activity ◽  
Granule Fraction ◽  
Alpha Beta

Collagenolytic cathepsin activity was detected in lysed rabbit peritoneal polymorphonuclear leucocytes. The pH optimum was around 3, and activity was greatly enhanced by the presence of cysteine and EDTA. Digestion of polymeric collagen resulted in the release of alpha, beta, and gamma-chains. Collagenolytic cathepsin activity was associated mainly with the granule fraction isolated from homogenates by differential centrifugation. The granule fraction was further fractionated by isopycnic density-gradient centrifugation, and the collagenolytic cathepsin activity was shown to be associated with the azurophil and tertiary granules, both lysosome-like organelles.

NATURE OF NEWLY SYNTHESIZED RNA IN DEVELOPING RAT BRAIN

1965 ◽  
Vol 43 (7) ◽  
pp. 1083-1089 ◽  
Author(s):  
Udai N. Singh
Keyword(s):  
Rat Brain ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Brain Slices ◽  
Sucrose Density Gradient ◽  
Major Fraction ◽  
Sucrose Density Gradient Centrifugation ◽  
Young Rat ◽  
Developing Rat

Sedimentation characteristics of labelled RNA synthesized in young rat brain slices have been studied by sucrose density gradient centrifugation. A major fraction of the radioactivity derived from adenine-8-C14 in phenol-extractable RNA is shown to be localized in soluble RNA and in relatively small amounts of highly labile fractions in the region lying between 10 S and 20 S. Smaller peaks with S values greater than 30 are also observed. The radioactivity profile of a rapidly labelled RNA isolated from interphase is found to be in sharp contrast with that of phenol-extractable RNA.

Uranyl Acetate and Rotary Shadowing to Increase the Contrast of Small Aggregated Virus Particles

1969 ◽  
Vol 27 ◽  
pp. 424-425
Author(s):  
Lee F. Ellis ◽  
Richard M. Van Frank ◽  
Walter J. Kleinschmidt
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Uranyl Acetate ◽  
Interferon Inducer ◽  
Virus Particles ◽  
Staining Methods ◽  
Rotary Shadowing

The extract from Penicillum stoliniferum, known as statolon, has been purified by density gradient centrifugation. These centrifuge fractions contained virus particles that are an interferon inducer in mice or in tissue culture. Highly purified preparations of these particles are difficult to enumerate by electron microscopy because of aggregation. Therefore a study of staining methods was undertaken.

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