Cell Biology

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    Protocols in Current Issue
    Fluorescent Labeling of Small Extracellular Vesicles (EVs) Isolated from Conditioned Media
    Authors:  John Santelices, Mark Ou, Winnie W. Hui, Gustavo H. B. Maegawa and Mariola J. Edelmann, date: 06/20/2022, view: 1524, Q&A: 0
    [Abstract]

    Extracellular vesicles (EVs), such as exosomes, are produced by all known eukaryotic cells, and constitute essential means of intercellular communication. Recent studies have unraveled the important roles of EVs in migrating to specific sites and cells. Functional studies of EVs using in vivo and in vitro systems require tracking these organelles

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    Immunoisolation of Endosomal Recycling Vesicles from Saccharomyces cerevisiae
    Authors:  Sho W. Suzuki and Scott D. Emr, date: 05/05/2022, view: 609, Q&A: 0
    [Abstract]

    Endosomal recycling is essential for the appropriate function of the endosome. During this process, endosomal coat complexes (i.e., retromer, and Mvp1) are recruited to the endosome, and deform its membrane to form recycling vesicles. To further analyze this, we developed a protocol for the immunoisolation of recycling vesicles from budding yeast.

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    Fractionation and Extraction of Crude Nuclear Proteins From Arabidopsis Seedlings
    Authors:  Jiajia Zhao, Jinsong Bao and Feifei Xu, date: 01/20/2022, view: 1327, Q&A: 0
    [Abstract]

    The plant nucleus is an important subcellular organelle that contains the genome, ribosomal RNA, and regulatory proteins, and performs a central role in the functioning and metabolism of the cell. Fractionation of intact nuclei is a crucial process to elucidate the function of nuclear proteins. Here, we present a simple method for the

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    Isolation and Characterization of Membrane Vesicles from Lactobacillus Species
    Authors:  Julie C. Caruana, Scott N. Dean and Scott A. Walper, date: 09/05/2021, view: 1891, Q&A: 0
    [Abstract]

    Throughout their life cycle, bacteria shed portions of their outermost membrane comprised of proteins, lipids, and a diversity of other biomolecules. These biological nanoparticles have been shown to have a range of highly diverse biological activities, including pathogenesis, community regulation, and cellular defense (among others). In recent

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    Protein Import Assay into Mitochondria Isolated from Human Cells
    Authors:  Lena M. Murschall, Esra Peker, Thomas MacVicar, Thomas Langer and Jan Riemer, date: 06/20/2021, view: 2700, Q&A: 0
    [Abstract]

    Mitochondria are essential organelles containing approximately 1,500 proteins. Only approximately 1% of these proteins are synthesized inside mitochondria, whereas the remaining 99% are synthesized as precursors on cytosolic ribosomes and imported into the organelle. Various tools and techniques to analyze the import process have been developed.

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    Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex
    Authors:  Ariel Diaz, Enrique Torre and Manuel Yepes, date: 01/20/2021, view: 2559, Q&A: 1
    [Abstract]

    The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the

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    Lipid Droplet Isolation from Arabidopsis thaliana Leaves
    Authors:  Yovanny Izquierdo, Rubén Fernández-Santos, Tomás Cascón and Carmen Castresana, date: 12/20/2020, view: 2282, Q&A: 0
    [Abstract]

    Lipid droplets (LDs) are neutral lipid aggregates surrounded by a phospholipid monolayer and specific proteins. In plants, they play a key role as energy source after seed germination, but are also formed in vegetative tissues in response to developmental or environmental conditions, where their functions are poorly understood. To elucidate these,

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    Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels
    Authors:  Eric Parigoris, Daniel L. Dunkelmann and Unai Silvan, date: 11/05/2020, view: 2800, Q&A: 0
    [Abstract] Giant unilamellar vesicles (GUVs) are a widely used model system for a range of applications including membrane biophysics, drug delivery, and the study of actin dynamics. While several protocols have been developed for their generation in recent years, the use of these techniques involving charged lipid types and buffers of physiological ionic ...
    Fluorescent Polysome Profiling in Caenorhabditis elegans
    Authors:  Dan Shaffer and Jarod A Rollins, date: 09/05/2020, view: 4104, Q&A: 0
    [Abstract] An important but often overlooked aspect of gene regulation occurs at the level of protein translation. Many genes are regulated not only by transcription but by their propensity to be recruited to actively translating ribosomes (polysomes). Polysome profiling allows for the separation of unbound 40S and 60S subunits, 80S monosomes, and actively ...
    Stopped-flow Light Scattering Analysis of Red Blood Cell Glycerol Permeability
    [Abstract] Stopped-Flow Light Scattering (SFLS) is a method devised to analyze the kinetics of fast chemical reactions that result in a significant change of the average molecular weight and/or in the shape of the reaction substrates. Several modifications of the original stopped-flow system have been made leading to a significant extension of its technical ...



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