Cell Biology


    Protocols in Current Issue
    Extraction and Quantification of Sphingolipids from Hemiptera Insects by Ultra-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry
    Authors:  Ni Wang, Xiaoxiao Shi, Chao Zhang, Wenwu Zhou and Zeng-Rong Zhu, date: 02/20/2021, view: 541, Q&A: 0

    Sphingolipids are major structural components of endomembranes and have also been described as an intracellular second messenger involved in various biological functions in all eukaryotes and a few prokaryotes. Ceramides (Cer), the central molecules of sphingolipids, have been depicted in cell growth arrest, cell differentiation, and apoptosis.

    Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages
    Authors:  Gretchen L. Seim, Steven V. John and Jing Fan, date: 07/20/2020, view: 1479, Q&A: 0
    [Abstract] Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular metabolism. Capturing a comprehensive picture of metabolism is vital for understanding the role of metabolic ...
    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: 1551, 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 ...
    Lipid-exchange Rate Assay for Lipid Droplet Fusion in Live Cells
    Authors:  Jia Wang, Boon Tin Chua, Peng Li and Feng-Jung Chen, date: 07/20/2019, view: 2880, Q&A: 0
    [Abstract] Lipid droplets (LDs) are central organelles in maintaining lipid homeostasis. Defective LD growth often results in the development of metabolic disorders. LD fusion and growth mediated by cell death–inducing DNA fragmentation factor alpha (DFFA)-like effector (CIDE) family proteins are crucial for various biological processes including unilocular ...
    Determining Oxidative Damage by Lipid Peroxidation Assay in Rat Serum
    Authors:  Qian Tang, Yu-Wen Su and Cory J. Xian, date: 06/20/2019, view: 4524, Q&A: 0
    [Abstract] It has been well-established that malondialdehyde (MDA), which is generated during the process of lipid peroxidation, is a commonly known biomarker for oxidative stress. Therefore, the serum levels of MDA are detected by using the lipid peroxidation assay with commercially available kit to determine the induction of oxidative stress in rat models.
    Fluorescence HPLC Analysis of the in-vivo Activity of Glucosylceramide Synthase
    Authors:  Kartik R. Roy, Sachin K. Khiste, Zhijun Liu and Yong-Yu Liu, date: 06/20/2019, view: 3236, Q&A: 0
    [Abstract] Almost all functions of cells or organs rely on the activities of cellular enzymes. Indeed, the in-vivo activities that directly represent the cellular effects of enzymes in live organs are critical importance to appreciate the roles enzymes play in modulating physiological or pathological processes, although assessments of such in-vivo ...
    Preparation of Lipid-Stripped Serum for the Study of Lipid Metabolism in Cell Culture
    Authors:  Aaron M. Hosios, Zhaoqi Li, Evan C. Lien and Matthew G. Vander Heiden, date: 06/05/2018, view: 9220, Q&A: 0
    [Abstract] Studying lipid metabolism in cultured cells is complicated by the fact that cells are typically cultured in the presence of animal serum, which contains a wide, variable, and undefined variety of lipid species. Lipid metabolism can impact cell physiology, signaling, and proliferation, and the ability to culture cells in the absence of exogenous ...
    Flow Cytometric Quantification of Fatty Acid Uptake by Mycobacterium tuberculosis in Macrophages
    Authors:  Evgeniya V. Nazarova, Maria Podinovskaia, David G. Russell and Brian C. VanderVen, date: 02/20/2018, view: 7681, Q&A: 0
    [Abstract] Mycobacterium tuberculosis (Mtb) has evolved to assimilate fatty acids from its host. However, until recently, there was no reliable way to quantify fatty acid uptake by the bacteria during host cell infection. Here we describe a new method to quantify fatty acid uptake by intracellular bacilli. We infect macrophages with Mtb ...
    Ex vivo Analysis of Lipolysis in Human Subcutaneous Adipose Tissue Explants
    Authors:  Pauline Decaunes, Anne Bouloumié, Mikael Ryden and Jean Galitzky, date: 02/05/2018, view: 5095, Q&A: 0
    [Abstract] Most studies of human adipose tissue (AT) metabolism and functionality have been performed in vitro on isolated mature adipocyte or in situ using the microdialysis technique (Lafontan, 2012). However, these approaches have several limitations. The use of mature isolated adipocytes is limiting as adipocytes are not in their ...
    Determination of the Glycolysis and Lipogenesis in Culture of Hepatocytes
    Authors:  Pierre-Damien Denechaud and Luis Fajas, date: 11/05/2016, view: 9331, Q&A: 0
    [Abstract] Metabolic flux analyses are needed to provide insights into metabolic regulation that occurs in cells. The current protocol describes fast and reproducible methods for determining glycolysis and de novo lipogenesis of hepatocytes. Primary culture of hepatocytes is an ‘in vitro’ model useful to study liver glucose and lipid ...

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