Protocols in Current Issue
    Biofilm Formation Assay in Pseudomonas syringae
    Authors:  Xiaolong Shao, Yingpeng Xie, Yingchao Zhang and Xin Deng, date: 05/20/2019, view: 239, Q&A: 0
    [Abstract] Pseudomonas syringae is a model plant pathogen that infects more than 50 plant species worldwide, thus leading to significant yield loss. Pseudomonas biofilm always adheres to the surfaces of medical devices or host cells, thereby contributing to infection. Biofilm formation can be visualized on numerous matrixes, including ...
    A Radioisotope-free Oligosaccharyltransferase Assay Method
    Authors:  Takahiro Yamasaki and Daisuke Kohda, date: 03/05/2019, view: 518, Q&A: 0
    [Abstract] Glycosylation of asparagine residues is widespread in Eukarya, and occurs in virtually all Archaea and some eubacterial species. A membrane-bound enzyme, oligosaccharyltransferase, catalyzes the transfer of an oligosaccharide chain from a sugar donor (lipid-linked oligosaccharide, LLO) to an asparagine residue in the consensus sequence, ...
    Determination of Storage (Starch/Glycogen) and Total Saccharides Content in Algae and Cyanobacteria by a Phenol-Sulfuric Acid Method
    Authors:  Tomáš Zavřel, Petra Očenášová, Maria A. Sinetova and Jan Červený, date: 08/05/2018, view: 1965, Q&A: 0
    [Abstract] This is a protocol for quantitative determination of storage and total carbohydrates in algae and cyanobacteria. The protocol is simple, fast and sensitive and it requires only few standard chemicals. Great advantage of this protocol is that both storage and total saccharides can be determined in the cellular pellets that were already used for ...
    Glycogen and Extracellular Glucose Estimation from Cyanobacteria Synechocystis sp. PCC 6803
    Authors:  Md. Rezaul Islam Khan, Yushu Wang, Shajia Afrin, Lin He and Gang Ma, date: 05/05/2018, view: 1758, Q&A: 0
    [Abstract] Cyanobacteria, which have the extraordinary ability to grow using sunlight and carbon dioxide, are emerging as a green host to produce value-added products. Exploitation of this highly promising host to make products may depend on the ability to modulate the glucose metabolic pathway; it is the key metabolic pathway that generates intermediates ...
    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: 2993, 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 Intracellular Osmolytes in Cyanobacterial Cells
    Authors:  Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 1663, Q&A: 0
    [Abstract] Most of the cyanobacteria accumulate osmolytes including sucrose, glucosylglycerol, in their cells in response to salt stress. Here we describe a protocol of our laboratory for extraction and quantification of cyanobacterial intracellular sucrose and glucosylglycerol. We have confirmed this protocol was applicable to at least four kinds of ...
    Laminarin Quantification in Microalgae with Enzymes from Marine Microbes
    Authors:  Stefan Becker and Jan-Hendrik Hehemann, date: 04/20/2018, view: 2123, Q&A: 0
    [Abstract] The marine beta-glucan laminarin is an abundant storage polysaccharide in microalgae. High production rates and rapid digestion by heterotrophic bacteria turn laminarin into an ideal carbon and energy source, and it is therefore a key player in the marine carbon cycle. As a main storage glucan laminarin also plays a central role in the energy ...
    Bacterial Cell Wall Precursor Phosphatase Assays Using Thin-layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC)
    Authors:  Manuel Pazos, Christian Otten and Waldemar Vollmer, date: 03/20/2018, view: 2024, Q&A: 0
    [Abstract] Peptidoglycan encases the bacterial cytoplasmic membrane to protect the cell from lysis due to the turgor. The final steps of peptidoglycan synthesis require a membrane-anchored substrate called lipid II, in which the peptidoglycan subunit is linked to the carrier lipid undecaprenol via a pyrophosphate moiety. Lipid II is the target of ...
    Preparation of Onion Epidermal Cell Walls for Imaging by Atomic Force Microscopy (AFM)
    Authors:  Tian Zhang and Daniel J. Cosgrove, date: 12/20/2017, view: 4143, Q&A: 0
    [Abstract] The growing plant cell wall is comprised of long, thin cellulose microfibrils embedded in a hydrated matrix of polysaccharides and glycoproteins. These components are typically constructed in layers (lamellae) on the inner surface of the cell wall, i.e., between the existing wall and the plasma membrane. The organization of these ...
    Preparation of the Partially Methylated Alditol Acetates Derived from CS Tetrasaccharides Containing Galactose for the Gas Chromatography/Mass Spectrometry Analysis
    Authors:  Kyohei Higashi and Toshihiko Toida, date: 11/05/2017, view: 3569, Q&A: 0
    [Abstract] Chondroitin sulfate (CS), a member of the glycosaminoglycan (GAG) family of carbohydrates, is composed of linear, sulfated repeating disaccharide sequences of N-acetyl-D-galactosamine (GalNAc) and glucuronic acid (GlcA). Recently, a keratan sulfate (KS) disaccharide [GlcNAc6S(β1-3)Galactose(β1-]-branched CS-E was identified ...

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