Cell Biology

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    Protocols in Current Issue
    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: 470, 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: 587, 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: 1221, 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: 3641, 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: 3791, 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: 2728, 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: 6926, 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 ...
    A Technique for the Measurement of in vitro Phospholipid Synthesis via Radioactive Labeling
    Authors:  Lucia Rodriguez-Berdini and Gabriel O. Ferrero, date: 01/20/2016, view: 7987, Q&A: 0
    [Abstract] This is an assay designed to examine the radioactive phosphorous incorporation when the molecule is being synthesized, which means that only de novo synthesized phospholipids can be detected. Thus, with this technique it is possible to detect in vitro phospholipid synthesis under different required experimental conditions respect ...
    Assessment of Brown Adipocyte Thermogenic Function by High-throughput Respirometry
    Authors:  Kiana Mahdaviani, Ilan Benador and Orian Shirihai, date: 11/05/2015, view: 8941, Q&A: 0
    [Abstract] Brown adipose tissue (BAT) has the unique ability to dramatically increase mitochondrial uncoupled fuel oxidation for thermogenesis in response to adrenergic stimulation. A key parameter in assessing brown adipocyte thermogenic capacity is mitochondrial uncoupling as determined by respiration. Measuring mitochondrial oxygen consumption rate (OCR) ...
    13C Kinetic Labeling and Extraction of Metabolites from Adherent Mammalian Cells
    Authors:  Parul Mehrotra, Najmuddin Saquib and Kanury V. S. Rao, date: 04/20/2015, view: 6523, Q&A: 0
    [Abstract] Fluctuations in metabolite levels in mammalian cells are the most direct form of readout of the cellular metabolic state. The current protocol describes a method for pulse labeling and subsequent isolation of metabolites from adherent mammalian cells. The isolated metabolites can be identified and quantified by mass-spectrometry, allowing for ...



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