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Centrifugation, Density Gradient: Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
 JoVE Developmental Biology

Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids

1Center for Biomedicine, European Academy Bozen/Bolzano (EURAC), 2Laboratory of Medical Genetics, Fondazione IRCCS Ca´ Granda, Ospedale Maggiore Policlinico, 3Del E. Webb Center for Neuroscience, Aging & Stem Cell Research, Sanford-Burnham Medical Research Institute


JoVE 52885

 JoVE Immunology and Infection

Isolation of Exosomes from the Plasma of HIV-1 Positive Individuals

1Department of Microbiology, Biochemistry, Immunology, Morehouse School of Medicine, 2Department of Medicine, Emory University School of Medicine, 3Department of Pathology and Laboratory Medicine, Emory University School of Medicine, 4Yerkes National Primate Research Center


JoVE 53495

 JoVE Biology

Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs

1Sinclair Centre for Regenerative Medicine, Ottawa Hospital Research Institute, 2University of Ottawa, 3Department of Neonatology and Pediatric Critical Care Medicine, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, 4DFG Research Center and Cluster of Excellence for Regenerative Therapies (CRTD), Technische Universität, Dresden, 5Children's Hospital of Eastern Ontario Research Institute


JoVE 53782

 JoVE Immunology and Infection

Isolation of Leukocytes from the Human Maternal-fetal Interface

1Perinatology Research Branch, NICHD/NIH/DHHS, 2Department of Obstetrics and Gynecology, University of Michigan, 3Department of Epidemiology and Biostatistics, Michigan State University, 4Department of Molecular Obstetrics and Genetics, Wayne State University, 5Department of Obstetrics and Gynecology, Wayne State University School of Medicine, 6Department of Immunology and Microbiology, Wayne State University School of Medicine


JoVE 52863

 JoVE Immunology and Infection

Promoting 3-D Aggregation of FACS Purified Thymic Epithelial Cells with EAK 16-II/EAKIIH6 Self-assembling Hydrogel

1Institute of Cellular Therapeutics, Allegheny Health Network, 2Division of Pharmaceutical Sciences, Mylan School of Pharmacy, Duquesne University, 3Department of Biological Sciences, Carnegie Mellon University


JoVE 54062

 JoVE Immunology and Infection

Reliable and High Efficiency Extraction of Kidney Immune Cells

1Division of Nephrology, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital, 2Division of Biomedical Sciences, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital, 3Division of Endocrinology, Department of Medicine, University of Missouri-Columbia and Harry S Truman Memorial Veteran's Hospital


JoVE 54368

 JoVE Immunology and Infection

Large-Scale Purification of Porcine or Bovine Photoreceptor Outer Segments for Phagocytosis Assays on Retinal Pigment Epithelial Cells

1INSERM, U968, 2Sorbonne Universités, UPMC Paris 06, UMR_S 968, Institut de la Vision, 3CNRS, UMR_7210, 4Department of Biological Sciences, Center for Cancer, Genetic Diseases and Gene Regulation, Fordham University


JoVE 52100

 JoVE Biology

Separation of Spermatogenic Cell Types Using STA-PUT Velocity Sedimentation

1Department of Cell and Developmental Biology, University of Pennsylvania, 2Biomedical Graduate Studies, University of Pennsylvania, 3Department of Animal Biology and Center for Animal Transgenesis and Germ Cell Research, University of Pennsylvania, 4Department of Genetics, University of Pennsylvania, 5Department of Biology, University of Pennsylvania


JoVE 50648

 JoVE In-Press

Induced Pluripotent Stem Cell Generation from Blood Cells Using Sendai Virus and Centrifugation

1CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 2Division of Rheumatology, Department of Internal Medicine, Seoul St. Marys Hospital, Institute of Medical Science, College of Medicine, The Catholic University of Korea

Video Coming Soon

JoVE 54650

 Science Education: Essentials of Biochemistry

Density Gradient Ultracentrifugation

JoVE Science Education

Density gradient ultracentrifugation is a common technique used to isolate and purify biomolecules and cell structures. This technique exploits the fact that, in suspension, particles that are more dense than the solvent will sediment, while those that are less dense will float. A high-speed ultracentrifuge is used to accelerate this process in order to separate biomolecules within a density gradient, which can be established by layering liquids of decreasing density in a centrifuge tube. The video will cover the principles of density gradient ultracentrifugation, including a procedure that demonstrates sample preparation, creation of a sucrose gradient, ultracentrifugation, and collection of fractionated analytes. The applications section discusses isolation of multi-protein complexes, isolation of nucleic acid complexes, and separation using cesium chloride density gradients. Density gradient ultracentrifugation is a common approach to isolate and purify cell structures for biochemical experiments. The technique uses a high-speed, or ultra, centrifuge to nondestructively separate cellular components in a density gradient. This video describes the principles of density gradient ultracentrifugation, provides a general procedure using a sucrose gradient, and discusses some applications.

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