Cancer Biology

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    Protocols in Current Issue
    Analysis of Leukemia Cell Metabolism through Stable Isotope Tracing in Mice
    Authors:  Nick van Gastel, Jessica B. Spinelli, Marcia C. Haigis and David T. Scadden, date: 10/05/2021, view: 1386, Q&A: 0
    [Abstract]

    Once thought to be a mere consequence of the state of a cell, intermediary metabolism is now recognized as a key regulator of mammalian cell fate and function. In addition, cell metabolism is often disturbed in malignancies such as cancer, and targeting metabolic pathways can provide new therapeutic options. Cell metabolism is mostly studied in

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    Monitoring Changes in the Oxidizing Milieu in the Endoplasmic Reticulum of Mammalian Cells Using HyPerER
    Authors:  Julia Birk, Beata Lizak, Christian Appenzeller-Herzog and Alex Odermatt, date: 07/05/2021, view: 1764, Q&A: 0
    [Abstract]

    The production of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress are tightly linked. The generation of ROS can be both the cause and a consequence of ER stress pathways, and an increasing number of human diseases are characterized by tissue atrophy in response to ER stress and oxidative injury. For the assessment of modulators

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    Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
    Authors:  Birte Plitzko and Sandra Loesgen, date: 05/20/2018, view: 60640, Q&A: 1
    [Abstract] Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual ...
    FACS-based Glucose Uptake Assay of Mouse Embryonic Fibroblasts and Breast Cancer Cells Using 2-NBDG Probe
    Authors:  Shengli Dong and Suresh K Alahari, date: 04/20/2018, view: 8295, Q&A: 0
    [Abstract] This is a flow cytometry-based protocol to measure glucose uptake of mouse embryonic fibroblasts (MEFs) and breast cancer cells in vitro. The method is a slightly modified and updated version as previously described (Dong et al., 2017). Briefly, the target cells are incubated with the fluorescently tagged ...
    Assessment of Cellular Redox State Using NAD(P)H Fluorescence Intensity and Lifetime
    Authors:  Thomas S. Blacker, Tunde Berecz, Michael R. Duchen and Gyorgy Szabadkai, date: 01/20/2017, view: 11001, Q&A: 0
    [Abstract] NADH and NADPH are redox cofactors, primarily involved in catabolic and anabolic metabolic processes respectively. In addition, NADPH plays an important role in cellular antioxidant defence. In live cells and tissues, the intensity of their spectrally-identical autofluorescence, termed NAD(P)H, can be used to probe the mitochondrial redox state, ...
    Fat Turnover Assay in Drosophila
    Authors:  Subhash D. Katewa and Pankaj Kapahi, date: 11/05/2016, view: 10276, Q&A: 0
    [Abstract] Like all animals, Drosophila shows robust fat (triglyceride) turnover, i.e., they synthesize, store and utilize triglyceride for their daily metabolic needs. The protocol describes a simple assay to measure this turnover of triglycerides in Drosophila.
    Spectrophotometric Determination of Glutamine Synthetase Activity in Cultured Cells
    Authors:  I-Chen Peng, Alex J. Bott and Wei-Xing Zong, date: 10/05/2016, view: 11291, Q&A: 0
    [Abstract] Glutamine synthetase (GS), which catalyzes the conversion of glutamate and ammonia to glutamine, is widely distributed in animal tissues and cell culture lines. The importance of this enzyme is suggested by the fact that glutamine, the product of GS-catalyzed de novo synthesis reaction, is the most abundant free amino acid in blood (Smith and ...
    Measurement of Transferrin- and Non-transferrin-bound Iron Uptake by Mouse Tissues
    Authors:  Supak Jenkitkasemwong, Chia-Yu Wang and Mitchell D. Knutson, date: 09/05/2016, view: 7345, Q&A: 0
    [Abstract] Iron in blood plasma is bound to its transport protein transferrin, which delivers iron to most tissues. In iron overload and certain pathological conditions, the carrying capacity of transferrin can become exceeded, giving rise to non-transferrin-bound iron, which is taken up preferentially by the liver, kidney, pancreas, and heart. The ...
    Quantification of 2-Hydroxyglutarate Enantiomers by Liquid Chromatography-mass Spectrometry
    Authors:  William M. Oldham and Joseph Loscalzo, date: 08/20/2016, view: 9382, Q&A: 0
    [Abstract] Two enantiomers of 2-hydroxyglutarate (2HG), L (L2HG) and D (D2HG), are metabolites of unknown function in mammalian cells that were initially associated with separate and rare inborn errors of metabolism resulting in increased urinary excretion of 2HG linked to neurological deficits in children (Chalmers et al., 1980; Duran et al ...
    Extraction and Quantification of Tryptophan and Kynurenine from Cultured Cells and Media Using a High Performance Liquid Chromatography (HPLC) System Equipped with an Ultra-sensitive Diode Array Detector
    Authors:  Jeffrey Kim, Benjamin Stewart and Robert H Weiss, date: 04/05/2016, view: 7317, Q&A: 0
    [Abstract] Evidence of the involvement of tryptophan and its metabolite, kynurenine, in various biological processes including cancer is constantly expanding. Analysis of cell extracts and culture media can allow for quick snapshots of the metabolic fluctuations occurring in vitro. Here, we describe a method for metabolite extraction from mammalian ...



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