Protocols in Current Issue
    Isolation of Tumor Cells Based on Their Distance from Blood Vessels
    Authors:  Saran Kumar, Husni Sharife, Tirzah Kreisel, Libat Bar-Lev, Myriam Grunewald and Eli Keshet, date: 05/20/2020, view: 871, Q&A: 0
    [Abstract] Differential exposure of tumor cells to microenvironmental cues greatly impacts cell phenotypes, raising a need for position based sorting of tumor cells amenable to multiple OMICs and functional analyses. One such key determinant of tumor heterogeneity in solid tumors is its vasculature. Proximity to blood vessels (BVs) profoundly affects tumor ...
    Single-run HPLC Quantification of Plant Cell Wall Monosaccharides
    [Abstract] The plant cell wall is a complex network of polysaccharides and proteins that provides strength and structural integrity to plant cells, as well as playing a vital role in growth, development, and defense response. Cell wall polysaccharides can be broadly grouped into three categories: cellulose, pectins, and hemicelluloses. Dynamic interactions ...
    Steady-state and Flux-based Trehalose Estimation as an Indicator of Carbon Flow from Gluconeogenesis or Glycolysis
    Authors:  Ritu Gupta and Sunil Laxman, date: 01/05/2020, view: 1260, Q&A: 0
    [Abstract] Trehalose (and glycogen) is a major storage carbohydrate in many cells, including S. cerevisiae. Typically, trehalose (a disaccharide of glucose) is synthesized and stored through gluconeogenesis. However, trehalose can also be made directly from glucose, if glucose-6-phosphate is channeled away from glycolysis or pentose phosphate ...
    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: 5440, 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 ...
    Determination of Intra- and Extracellular Glucose in Mycelium of Fusarium oxysporum
    Authors:  Carmen Ruiz-Roldan and M. Isabel G. Roncero, date: 07/20/2016, view: 7203, Q&A: 0
    [Abstract] To study alterations in the metabolism and/or in the transport of glucose during Fusarium oxysporum vegetative growth, we determined intracellular glucose levels in different fungal strains, as well as the amount of glucose remaining in the supernatants after growth in synthetic medium (SM) supplemented with either 0.05 or 2.5% glucose. ...
    13C Tracer Studies of Metabolism in Mouse Tumor Xenografts
    Authors:  Andrew N. Lane, Jun Yan and Teresa W-M. Fan, date: 11/20/2015, view: 8161, Q&A: 0
    [Abstract] Mice are widely used for human tumor xenograft studies of cancer development and drug efficacy and toxicity. Stable isotope tracing coupled with metabolomic analysis is an emerging approach for assaying metabolic network activity. In mouse models there are several routes of tracer introduction, which have particular advantages and disadvantages ...
    Analysis of Malondialdehyde, Chlorophyll Proline, Soluble Sugar, and Glutathione Content in Arabidopsis seedling
    Authors:  Zhijin Zhang and Rongfeng Huang, date: 07/20/2013, view: 24548, Q&A: 2
    [Abstract] The protocol has four sub-protocols, which are about the measurement of malondialdehyde, chlorophyll proline, soluble sugar, and glutathione content, respectively, in Arabidopsis seedling by using spectrophotometer. These methods are simple, effective and reproducible, which will help the researchers who are not familiar with these ...
    Glucose Production Assay in Primary Mouse Hepatocytes
    Authors:  Michihiro Matsumoto and Mashito Sakai, date: 11/05/2012, view: 21009, Q&A: 3
    [Abstract] Hepatic glucose production is a primary determinant of fasting hyperglycemia in type 2 diabetic patients. Glucagon-cAMP-PKA pathway increases, but insulin-PI3 kinase-Akt pathway suppresses glucose production. This assay aims to evaluate the ability of isolated mouse hepatocytes to release newly synthesized glucose mainly from lactate and pyruvate ...
    5-Hydroxymethylcytosine (5-hmC) Specific Enrichment
    Authors:  Keith E Szulwach, Chun-Xiao Song, Chuan He and Peng Jin, date: 08/05/2012, view: 8099, Q&A: 0
    [Abstract] 5-Hydroxymethylcytosine (5-hmC) is a newly discovered DNA modification in mammalian genomes. This protocol is to be a highly efficient and selective chemical approach to label and capture 5-hmC, taking advantage of a bacteriophage enzyme that adds a glucose moiety to 5-hmC specifically, which could in turn be used for high-throughput mapping via ...

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