Protocols in Current Issue
    Triacylglycerol Measurement in HeLa Cells
    Authors:  Ximing Du and Hongyuan Yang, date: 12/20/2020, view: 724, Q&A: 0

    Lipid droplets store triacylglycerols (triglycerides) and sterol esters to regulate lipid and energy homeostasis. Triacylglycerol measurement is often performed during the investigation of lipid droplet formation and growth. This protocol describes a reliable method using a fluorometric lipid quantification kit to measure triacylglycerols

    Total Triglyceride Quantification in Caenorhabditis elegans
    Authors:  Anjali Sandhu and Varsha Singh, date: 11/20/2020, view: 903, Q&A: 0
    [Abstract] Several studies suggest an important role of lipid metabolism in regulating longevity of Caenorhabditis elegans. Therefore, assays to quantify lipids have enormous value in understanding aging and pathologies associated with it. Approximately 70% of lipid metabolism genes in the nematode have orthologs in humans. Amenability of C. elegans ...
    Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels
    Authors:  Eric Parigoris, Daniel L. Dunkelmann and Unai Silvan, date: 11/05/2020, view: 1019, 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 ...
    Calcein Release Assay to Measure Membrane Permeabilization by Recombinant Alpha-Synuclein
    Authors:  Sayan Dutta, Ben G. Watson, Seema Mattoo and Jean-Christophe Rochet, date: 07/20/2020, view: 1281, Q&A: 0
    [Abstract] Lipid membranes are involved in regulating biochemical and biological processes and in modulating the selective permeability of cells, organelles, and vesicles. Membrane composition, charge, curvature, and fluidity all have concerted effects on cellular signaling and homeostasis. The ability to prepare artificial lipid assemblies that mimic ...
    Isolation of Lipid Rafts from Cultured Mammalian Cells and Their Lipidomics Analysis
    Authors:  Nigora Mukhamedova, Kevin Huynh, Hann Low, Peter J. Meikle and Dmitri Sviridov, date: 07/05/2020, view: 1332, Q&A: 0
    [Abstract] Lipid rafts are distinct liquid-ordered domains of plasma membranes of most eukaryotic cells providing platform for signaling pathways. Lipid composition of rafts is critical for their structural integrity and for regulation of signaling pathways originating from rafts. Here we provide a protocol to isolate lipid rafts from cultured human and ...
    Quantification of Fatty Acids in Mammalian Tissues by Gas Chromatography–Hydrogen Flame Ionization Detection
    [Abstract] In mammalian organisms, fatty acids (FAs) exist mostly in esterified forms, as building blocks of phospholipids, triglycerides, and cholesteryl esters, while some exist as non-esterified free FAs. The absolute quantification of FA species in total lipids or in a specific lipid class is critical in lipid-metabolism studies. To quantify FAs in ...
    Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids
    Authors:  Shunshi Kohyama, Kei Fujiwara, Natsuhiko Yoshinaga and Nobuhide Doi, date: 03/20/2020, view: 1658, Q&A: 0
    [Abstract] The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling ...
    In vitro Measurement of CMP-Sialic Acid Transporter Activity in Reconstituted Proteoliposomes
    Authors:  James Cahill, Shivani Ahuja and Matthew R. Whorton, date: 03/20/2020, view: 1422, Q&A: 0
    [Abstract] Nucleotide-sugar transporters (NSTs) facilitate eukaryotic cellular glycosylation by transporting nucleotide-sugar conjugates into the Golgi lumen and endoplasmic reticulum for use by glycosyltransferases, while also transferring nucleotide monophosphate byproducts to the cytoplasm. Mutations in this family of proteins can cause a number of ...
    Contemporaneous Measurement of Outer and Inner Membrane Permeability in Gram-negative Bacteria
    Authors:  Bo Ma, Chao Fang, Jing Zhang, Mingzhi Wang, Xiaoxing Luo and Zheng Hou, date: 03/05/2020, view: 1545, Q&A: 0
    [Abstract] The emergence and rapid spread of multidrug resistance in bacteria have led to the urgent need for novel antibacterial agents. Membrane permeabilization is the mechanism for many antibacterial molecules that are being developed against gram-negative bacteria. Thus, to determine the efficacy of a potential antibacterial molecule, it is important to ...
    Lipid Mixing Assay for Murine Myoblast Fusion and Other Slow Cell-cell Fusion Processes
    [Abstract] Lipid mixing (redistribution of lipid probes between fusing membranes) has been widely used to study early stages of relatively fast viral and intracellular fusion processes that take seconds to minutes. Lipid mixing assays are especially important for identification of hemifusion intermediates operationally defined as lipid mixing without content ...

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