Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

10.3791/62119 ◽  
2021 ◽  
Author(s):  
William McCaig ◽  
Timothy LaRocca
Keyword(s):  
Density Gradient ◽  
Cell Fractionation ◽  
U937 Cells ◽  
Density Gradient Purification

Dextran Density Gradient Purification of Plasmodium Sporozoites

1987 ◽  
Vol 73 (6) ◽  
pp. 1261
Author(s):  
Peter Goldie ◽  
Michael J. Stewart ◽  
Seymour Schulman ◽  
Jerome P. Vanderberg
Keyword(s):  
Density Gradient ◽  
Density Gradient Purification

scholarly journals Preparation of Crude Subcellular Fractions by Differential Centrifugation

2002 ◽  
Vol 2 ◽  
pp. 1638-1642 ◽  
Author(s):  
John Graham
Keyword(s):  
Density Gradient ◽  
Buoyant Density ◽  
Mitochondrial Fraction ◽  
Subcellular Fractions ◽  
Low Density ◽  
Differential Centrifugation ◽  
Density Gradient Purification

The employment of differential centrifugation to prepare crude fractions of subcellular particles from homogenates is often a necessary first step to a subsequent purification of one or more particles on a density gradient. Buoyant density gradient purification of peroxisomes or lysosomes for example is almost invariably carried out on a light mitochondrial fraction so as to eliminate smaller particles that may have similar densities. Unless they are first removed, large rapidly sedimenting particles in homogenates may also disturb shallow gradients designed to fractionate small low-density microsomes.

The subcellular distribution and properties of Crithidia sp. hydrolases with particular reference to pyrophosphate and orthophosphate monoester phosphohydrolases

1976 ◽  
Vol 54 (4) ◽  
pp. 365-381 ◽  
Author(s):  
J. McLaughlin ◽  
H. S. Injeyan ◽  
E. Meerovitch
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Hydrolytic Enzymes ◽  
Hydrolytic Activity ◽  
Cell Fractionation ◽  
Acid Hydrolases ◽  
Mitochondrial Marker ◽  
Fractionation Scheme ◽  
A Cell

A cell fractionation scheme was developed for studying the distribution of certain hydrolases, especially phosphohydrolases, in a Crithidia sp. (Trypanosomatidae). Whilst between 26–56% of the total cellular hydrolase activities were soluble (probably of flagellar pocket origin), a certain percentage, 5–40%, was sedimentable. A particulate fraction obtained after isopycnic density gradient centrifugation (ρ = 1.187–1.241), designated fraction FA/FB, was enriched in various acid hydrolases (relative specific activities 1.33–6.24) and displayed latent phosphohydrolase activities. The density gradient distributions of these hydrolytic enzymes were compared with reference to one another and malate dehydrogenase (mitochondrial marker). From the results obtained it appears that the sedimentable acid hydrolases of Crithidia are associated with a heterogeneous population of subcellular particles. Cytochemical observations on the FA/FB fraction supported this finding and revealed the association of acid phosphatase reaction product with subcellular elements resembling multivesicular bodies.A detailed study of the hydrolytic activity toward both ortho- and pyrophosphate substrates by the soluble, as compared to the FA/FB fractions, revealed the existence of distinct phosphohydrolase activities. It is proposed that these differences in properties are due to the existence of phosphohydrolase isoenzymes and that they in turn relate to a differentiation in the cellular digestive functions of Crithidia.

Immunobead and Density Gradient Purification of Paget Cells:In vitroStudies of Proliferation

1998 ◽  
Vol 25 (8) ◽  
pp. 497-502 ◽  
Author(s):  
Osamu Mori ◽  
Minoru Miyasato ◽  
Tadashi Karashima ◽  
Takashi Hashimoto
Keyword(s):  
Density Gradient ◽  
Density Gradient Purification

scholarly journals Density Gradient Purification of Mononuclear Cells Is Associated with Spurious (Artifactual) Immunosuppression

1988 ◽  
Vol 208 (6) ◽  
pp. 768-775 ◽  
Author(s):  
DAZHONG XU ◽  
EDWIN A. DEITCH ◽  
KEVIN SITTIG ◽  
L U QI ◽  
JOHN C. MCDONALD
Keyword(s):  
Density Gradient ◽  
Mononuclear Cells ◽  
Density Gradient Purification

scholarly journals A rapid and scalable density gradient purification method for Plasmodium sporozoites

2012 ◽  
Vol 11 (1) ◽  
pp. 421 ◽  
Author(s):  
Mark Kennedy ◽  
Matthew E Fishbaugher ◽  
Ashley M Vaughan ◽  
Rapatbhorn Patrapuvich ◽  
Rachasak Boonhok ◽  
...  
Keyword(s):  
Density Gradient ◽  
Purification Method ◽  
Density Gradient Purification

scholarly journals Pancreatic islet isolation by mechanical-enzymatic separation, stationary collagenase digestion and dextran discontinuous density gradient purification: experimental study in dogs

2002 ◽  
Vol 17 (2) ◽  
pp. 89-94 ◽  
Author(s):  
Jaques Waisberg ◽  
Heloísa Prado Soares ◽  
Luís Gustavo Altieri ◽  
Osíris Ramaciotti ◽  
Manlio Basilio Speranzini
Keyword(s):  
Experimental Study ◽  
Body Weight ◽  
Pancreatic Islets ◽  
Density Gradient ◽  
Gradient Methods ◽  
Islet Isolation ◽  
Single Donor ◽  
Collagenase Digestion ◽  
Pancreatic Islet Isolation ◽  
Density Gradient Purification

The prospects for allotransplantation of pancreatic islets in man depend on the development of methods that provide sufficient quantities of pancreatic islets from a single donor, which are capable, when transplanted, of achieve the normalization of carbohydrate metabolism. Objective: Evaluate the efficacy of the isolation of Langerhans islets from dogs, by means of mechanical-enzymatic separation technique with stationary digestion using collagenase, and purification with a discontinuous dextran density gradient. Methods: The counting of islet numbers and evaluation of their sizes was accomplished by staining with diphenylthiocarbazone and using stereoscopic microscopes equipped with eyepiece reticule for the measurement of average diameters of stained islets. Results: The results disclosed that the average number of islets isolated was 81032.20 ± 24736.79 and the average number of islets isolated per kg of body weight was 6938.70 ± 1392.43. The average number of islets isolated per kg of body weight showed significant correlation with body weight and weight of the pancreas resected. Conclusion: The number of islets isolated, of a single donor, by mechanical-enzymatic separation, stationary collagenase digestion and discontinuous dextran density gradient purification can be sufficient to success of pancreatic islets transplant in dogs.

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