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  JoVE General

  
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General Laboratory Techniques

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Basic Methods in Cellular and Molecular Biology

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Model Organisms I

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 JoVE General

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

The STA-PUT method allows for the separation of different populations of spermatogenic cells based on size and density.

 JoVE General

Assembly of Nucleosomal Arrays from Recombinant Core Histones and Nucleosome Positioning DNA

1Biochemistry and Molecular Biology, Colorado State University


JoVE 50354

A method is presented for the reconstitution of model nucleosomal arrays from recombinant core histones and tandemly repeated nucleosome positioning DNA. We also describe how sedimentation velocity experiments in the analytical ultracentrifuge, and atomic force microscopy (AFM) are used to monitor the extent of nucleosomal array saturation after reconstitution.

 JoVE General

FtsZ Polymerization Assays: Simple Protocols and Considerations

1Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen


JoVE 50844

Polymerization of FtsZ is essential for bacterial cell division. In this report, we detail simple protocols to monitor FtsZ polymerization activity and discuss the influence of buffer composition. The protocols can be used to study the interaction of FtsZ with regulatory proteins or antibacterial drugs that affect FtsZ polymerization.

 JoVE General

Isolation and Purification of Kinesin from Drosophila Embryos

1Department of Developmental and Cell Biology, School of Biosciences, University of California, Irvine


JoVE 3501

This is a protocol to isolate active full length Kinesin from Drosophila embryos for single-molecule biophysical studies. We show how to collect embryos, make the embryo lysate, and then polymerize microtubules (MTs). Kinesin is purified by immobilizing it on the MTs, spinning down the Kinesin-MT complexes, and then releasing the kinesin from the MTs via ATP addition.

 JoVE General

Assembly, Loading, and Alignment of an Analytical Ultracentrifuge Sample Cell

1National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Dynamics of Macromolecular Assembly, Laboratory of Bioengineering and Physical Science


JoVE 1530

The analytical ultracentrifuge (AUC) sample cell holds sample and reference buffer and during experiments and is exposed to high vacuum and rotor speeds up to 60,000 rpm. This video will demonstrate the rigorous attention to detail necessary for assembly, loading and alignment of this very important component of an AUC experiment.

 JoVE Bioengineering

Micro-particle Image Velocimetry for Velocity Profile Measurements of Micro Blood Flows

1Department of Chemical and Biological Engineering, University of Ottawa, 2Department of Mechanical Engineering, University of Ottawa


JoVE 50314

Micro-particle image velocimetry (μPIV) is used to visualize paired images of micro particles seeded in blood flows which are cross-correlated to give an accurate velocity profile. Shear rate, maximum velocity, velocity profile shape, and flow rate, each of which has clinical applications, can be derived from these measurements.

 JoVE Applied Physics

Fast Imaging Technique to Study Drop Impact Dynamics of Non-Newtonian Fluids

1Department of Physics, The University of Chicago, 2James Franck Institute, The University of Chicago, 3Department of Mechanical Engineering and Materials Science, Yale University


JoVE 51249

Drop impact of non-Newtonian fluids is a complex process since different physical parameters influence the dynamics over a very short time (less than one tenth of a millisecond). A fast imaging technique is introduced in order to characterize the impact behaviors of different non-Newtonian fluids.

 JoVE General

Simple and Efficient Technique for the Preparation of Testicular Cell Suspensions

1Departamento de Biología Molecular, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), 2Servicio de Citometría de Flujo y Clasificación Celular (SECIF), Instituto de Investigaciones Biológicas Clemente Estable, 3Sección Bioquímica, Facultad de Ciencias, Universidad de la República


JoVE 50102

A novel protocol for the mechanical preparation of testicular cell suspensions from rodent material, avoiding enzymes and detergents, is described. The method is very simple, fast, reproducible, and renders good quality cell suspensions, which are suitable for flow sorting and RNA extraction.

 JoVE Clinical and Translational Medicine

Isolation of Soluble and Insoluble PrP Oligomers in the Normal Human Brain

1Department of Pathology, National Prion Disease Pathology Surveillance Center, Case Western Reserve University School of Medicine, 2Department of Neurology, National Prion Disease Pathology Surveillance Center, Case Western Reserve University School of Medicine


JoVE 3788

A new species of cellular prion protein (PrPC) has recently been identified in uninfected human brains using the methods described here. These methods can be used to isolate various PrP species, while some of them are also useful in isolating other misfolded protein aggregates from human brains.

 JoVE General

Flow Cytometry Purification of Mouse Meiotic Cells

1Genome Plasticity Laboratory, Department of Cancer Biology, The Scripps Research Institute, 2Flow Cytometry Core, The Scripps Research Institute


JoVE 2602

An efficient method to obtain highly purified viable meiotic fractions from mouse testis is described, which combines a refined cell dissociation protocol with fluorescent activated cell sorting (FACS). This method takes advantage of differences in the DNA content and nuclear density of discrete meiotic fractions.

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