Density Gradient Centrifugation of Red Blood cells carrying the Mammary Tumour Virus

One step centrifugation procedure used commonly for separation of blood cells is the ficoll gradient centrifugation. In this method, after centrifugation, the peripheral blood mononuclear cells (PBMCs) are located on the top of the separation fluid, whereas other blood cells erythrocytes and granulocytes sediment to the bottom. In the present study 75% of lymphocyte suspension could be separated by using a one-step density gradient centrifugation of sodium heparin blood with Sucrose. Sucrose was diluted into different concentrations using miliQ water (10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 100%,). 4 mL of diluted blood was layered on 4 mL of each sucrose solution and centrifuged for 45 minutes at 1000 rpm. Clear separation of PBMCs could be observed in solution with 40% sucrose. The separated PBMCs were analysed in haeme analyser which showed 75% lymphocytes, 23% monocytes and 2% of other cells.

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The Nature of Monkey Cell Haemagglutinins of Rosen’s Group II Adenoviruses

Zeitschrift für Naturforschung B ◽

10.1515/znb-1965-0613 ◽

1965 ◽

Vol 20 (6) ◽

pp. 560-563 ◽

Cited By ~ 9

Author(s):

R. Wigand ◽

M. Stöhr

Keyword(s):

Density Gradient ◽

Blood Cells ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Buoyant Density ◽

Haemagglutination Inhibition ◽

Wild Strain ◽

Rat Blood ◽

Monkey Cell ◽

Group Ii

Some of the adenovirus type of Rosen’s group II exhibit weak agglutinating properties for monkey blood cells. The relation of these haemagglutinins to haemagglutinins for rat blood cells and to the infectious virus was studied by cross-absorption with blood cells and by density gradient centrifugation with caesium chloride. In the virus types studied (type 9, 13, and a wild strain of type 15) one part of the monkey cell haemagglutinin was associated with the virion and, for type 9 and 15, was inseparable from the virus-associated haemagglutinin for rat blood cells. In density gradient centrifugation a second haemagglutinin for monkey blood cells was found, separable from the virus particles and associated with the portion of higher buoyant density of the soluble haemagglutinin for rat blood cells. In type 13 and 23, haemagglutinins for monkey and rat blood cells can be separated by absorption with the heterologous blood cell species.Haemagglutinins for monkey cells are resistant to trypsin or trypsin plus subsequent heating to 55°C, in contrast to haemagglutinins for rat blood cells. Monkey cell haemagglutination is reversibly inhibited by the presence of CsCl.In haemagglutination-inhibition tests it was found that haemagglutination with monkey blood cells is more difficult to be inhibited by immune serum that haemagglutination with rat blood cells.

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Isolation of highly acidic and vanadium-containing blood cells from among several types of blood cell from ascidiidae species by density-gradient centrifugation

Journal of Experimental Zoology ◽

10.1002/jez.1402570304 ◽

1991 ◽

Vol 257 (3) ◽

pp. 306-313 ◽

Cited By ~ 73

Author(s):

Hitoshi Michibata ◽

Yukinari Iwata ◽

Junko Hirata

Keyword(s):

Blood Cell ◽

Density Gradient ◽

Blood Cells ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation

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Separation of viable microfilariae free of blood cells on Percoll gradients

Journal of Helminthology ◽

10.1017/s0022149x00028078 ◽

1984 ◽

Vol 58 (1) ◽

pp. 69-70 ◽

Cited By ~ 47

Author(s):

R. Chandrashekar ◽

U. R. Rao ◽

G. R. Rajasekariah ◽

D. Subrahmanyam

Keyword(s):

Density Gradient ◽

Blood Cells ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Brugia Pahangi ◽

Percoll Gradients ◽

Reproducible Method ◽

Dipetalonema Viteae

AbstractA consistent and reproducible method is described for isolating pure populations of microfilariae of Litomosoides carinii, Brugia pahangi, B. malayi and Dipetalonema viteae, free of cells, from blood, by density gradient centrifugation on Percoll in 0.25M sucrose. The recovery of the microfilariae was 85 to 97%.

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100064037 ◽

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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The Combining Ability of Glycoproteins IIb, IIIa and Ca2+ in EDTA-Treated Nonaggregable Platelets

Thrombosis and Haemostasis ◽

10.1055/s-0038-1665326 ◽

1983 ◽

Vol 50 (04) ◽

pp. 848-851 ◽

Cited By ~ 11

Author(s):

Marjorie B Zucker ◽

David Varon ◽

Nicholas C Masiello ◽

Simon Karpatkin

Keyword(s):

Density Gradient ◽

Combining Ability ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Electrophoretic Pattern ◽

Sucrose Density Gradient ◽

Irreversible Loss ◽

Sucrose Density Gradient Centrifugation ◽

Crossed Immunoelectrophoresis ◽

Triton X

SummaryPlatelets deprived of calcium and incubated at 37° C for 10 min lose their ability to bind fibrinogen or aggregate with ADP when adequate concentrations of calcium are restored. Since the calcium complex of glycoproteins (GP) IIb and IIIa is the presumed receptor for fibrinogen, it seemed appropriate to examine the behavior of these glycoproteins in incubated non-aggregable platelets. No differences were noted in the electrophoretic pattern of nonaggregable EDTA-treated and aggregable control CaEDTA-treated platelets when SDS gels of Triton X- 114 fractions were stained with silver. GP IIb and IIIa were extracted from either nonaggregable EDTA-treated platelets or aggregable control platelets with calcium-Tris-Triton buffer and subjected to sucrose density gradient centrifugation or crossed immunoelectrophoresis. With both types of platelets, these glycoproteins formed a complex in the presence of calcium. If the glycoproteins were extracted with EDTA-Tris-Triton buffer, or if Triton-solubilized platelet membranes were incubated with EGTA at 37° C for 30 min, GP IIb and IIIa were unable to form a complex in the presence of calcium. We conclude that inability of extracted GP IIb and IIIa to combine in the presence of calcium is not responsible for the irreversible loss of aggregability that occurs when whole platelets are incubated with EDTA at 37° C.

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