Protocols in Current Issue
    Extraction and Quantification of Polyphosphate (polyP) from Gram-negative Bacteria
    Authors:  Jan-Ulrik Dahl, Lihan Xie and Ursula Jakob, date: 09/20/2018, view: 3226, Q&A: 0
    [Abstract] Polyphosphate (polyP), a universally conserved biomolecule, is composed of up to 1,000 phosphate monomers linked via phosphoanhydride bonds. Reaching levels in bacteria that are in the high nmoles per mg protein range, polyP plays important roles in biofilm formation and colonization, general stress protection and virulence. Various protocols for ...
    Quantification of Hydrogen Sulfide and Cysteine Excreted by Bacterial Cells
    Authors:  Sergey Korshunov and James A. Imlay, date: 05/20/2018, view: 3751, Q&A: 0
    [Abstract] Bacteria release cysteine to moderate the size of their intracellular pools. They can also evolve hydrogen sulfide, either through dissimilatory reduction of oxidized forms of sulfur or through the deliberate or inadvertent degradation of intracellular cysteine. These processes can have important consequences upon microbial communities, because ...
    ROS Detection in Botryococcus braunii Colonies with CellROX Green Reagent
    [Abstract] We analyzed the reactive oxygen species (ROS) accumulation in the colony-forming green microalga Botryococcus braunii in response to several stress inducers such as NaCl, NaHCO3, salicylic acid (SA), methyl jasmonate, and acetic acid. A staining assay using the fluorescent dye CellROX Green was used. CellROX Green is a ...
    Heavy Metal Stress Assay of Caenorhabditis elegans
    Authors:  Strahil Iv. Pastuhov, Tatsuhiro Shimizu and Naoki Hisamoto, date: 06/05/2017, view: 6951, Q&A: 0
    [Abstract] Organisms have developed many protective systems to reduce the toxicity from heavy metals. The nematode Caenorhabditis elegans has been widely used to determine the protective mechanisms against heavy metals. Responses against heavy metals can be monitored by expression of reporter genes, while sensitivity can be determined by quantifying ...
    Highly Accurate Real-time Measurement of Rapid Hydrogen-peroxide Dynamics in Fungi
    Authors:  Michael Mentges and Jörg Bormann, date: 12/20/2016, view: 6740, Q&A: 0
    [Abstract] Reactive oxygen species (ROS) are unavoidable by-products of aerobic metabolism. Despite beneficial aspects as a signaling molecule, ROS are principally recognized as harmful agents that act on nucleic acids, proteins and lipids. Reactive oxygen species, and, in particular, hydrogen peroxide (H2O2), are deployed as defense ...
    Determination of (p)ppGpp Levels During Stringent Response in Streptomyces coelicolor by Thin Layer Chromatography
    Authors:  Smitha Sivapragasam and Anne Grove, date: 11/05/2016, view: 7517, Q&A: 0
    [Abstract] The stringent response in bacteria is a stress response that is mediated by the signaling molecules guanosine tetraphosphate and pentaphosphate [(p)ppGpp], alarmones that are also directly related to virulence. Therefore, determination of (p)ppGpp levels is crucial for studying the stringent response. The protocol here outlines in a step-wise ...

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