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    Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA
    Authors:  Christian Reuter and Tobias A. Oelschlaeger, date: 06/20/2018, view: 126, Q&A: 0
    The mucosal surfaces of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye harbor a resident microflora that lives in symbiosis with their host and forms a complex ecosystem. The protection of the ...
    Preparation of Cell-free Synthesized Proteins Selectively Double Labeled for Single-molecule FRET Studies
    Authors:  Mayuri Sadoine, Michele Cerminara, Jörg Fitter, Alexandros Katranidis and Rajneesh, date: 06/20/2018, view: 112, Q&A: 0
    Single-molecule FRET (smFRET) is a powerful tool to investigate molecular structures and conformational changes of biological molecules. The technique requires protein samples that are site-specifically equipped with a pair of donor and acceptor ...
    A Procedure for Precise Determination of Glutathione Produced by Saccharomyces cerevisiae
    Authors:  Jyumpei Kobayashi, Daisuke Sasaki, Akihiko Kondo and Rajneesh, date: 06/20/2018, view: 80, Q&A: 0
    In bioproduction, yields of products must be calculated precisely for accurate evaluation of various fermentation conditions. To evaluate productivity of microorganisms, product amounts per unit of medium volume (e.g., mg-product/L-broth), ...
    Analysis of Metals in Whole Cells, Thylakoids and Photosynthetic Protein Complexes in Synechocystis sp. PCC6803
    Authors:  Chiara Gandini, Søren Husted, Sidsel Birkelund Schmidt and Rajneesh, date: 06/20/2018, view: 85, Q&A: 0
    Metals are essential in many biological processes, including oxygenic photosynthesis. Here we described a method to measure the metal pool in whole cells and thylakoids, including the bioactive pool in intact photosynthetic protein complexes in the ...
    Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA
    Authors:  Christian Reuter and Tobias A. Oelschlaeger, date: 06/20/2018, view: 126, Q&A: 0
    [Abstract] The mucosal surfaces of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye harbor a resident microflora that lives in symbiosis with their host and forms a complex ecosystem. The protection of the vulnerable epithelium is primarily achieved by mucins that form a gel-like structure adherent to the apical ...
    Analysis of Metals in Whole Cells, Thylakoids and Photosynthetic Protein Complexes in Synechocystis sp. PCC6803
    Authors:  Chiara Gandini, Søren Husted, Sidsel Birkelund Schmidt and Rajneesh, date: 06/20/2018, view: 85, Q&A: 0
    [Abstract] Metals are essential in many biological processes, including oxygenic photosynthesis. Here we described a method to measure the metal pool in whole cells and thylakoids, including the bioactive pool in intact photosynthetic protein complexes in the model oxygenic cyanobacterium Synechocystis PCC6803. In the first part of the protocol, ...
    A Procedure for Precise Determination of Glutathione Produced by Saccharomyces cerevisiae
    Authors:  Jyumpei Kobayashi, Daisuke Sasaki, Akihiko Kondo and Rajneesh, date: 06/20/2018, view: 80, Q&A: 0
    [Abstract] In bioproduction, yields of products must be calculated precisely for accurate evaluation of various fermentation conditions. To evaluate productivity of microorganisms, product amounts per unit of medium volume (e.g., mg-product/L-broth), and/or product amounts per unit of a microorganism amount (e.g., mg-product/mg-dry cell ...
    Preparation of Cell-free Synthesized Proteins Selectively Double Labeled for Single-molecule FRET Studies
    Authors:  Mayuri Sadoine, Michele Cerminara, Jörg Fitter, Alexandros Katranidis and Rajneesh, date: 06/20/2018, view: 112, Q&A: 0
    [Abstract] Single-molecule FRET (smFRET) is a powerful tool to investigate molecular structures and conformational changes of biological molecules. The technique requires protein samples that are site-specifically equipped with a pair of donor and acceptor fluorophores. Here, we present a detailed protocol for preparing double-labeled proteins for smFRET ...
    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: 370, 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 ...
    Protocol for RYMV Inoculation and Resistance Evaluation in Rice Seedlings
    Authors:  Agnès Pinel-Galzi, Eugénie Hébrard, Oumar Traoré, Drissa Silué and Laurence Albar, date: 06/05/2018, view: 228, Q&A: 0
    [Abstract] Rice yellow mottle virus (RYMV), a mechanically transmitted virus that causes serious damage to cultivated rice plants, is endemic to Africa. Varietal selection for resistance is considered to be the most effective and sustainable management strategy. Standardized resistance evaluation procedures are required for the identification and ...
    Characterization of Protein Domain Function via in vitro DNA Shuffling
    Authors:  Kathy Hiu Laam Po, Edward Wai Chi Chan and Sheng Chen, date: 06/05/2018, view: 164, Q&A: 0
    [Abstract] We recently investigated the molecular events that drive evolution of the CTX-M-type β-lactamases by DNA shuffling of fragments of the blaCTX-M-14 and blaCTX-M-15 genes. Analysis of a total of 51 hybrid enzymes showed that enzymatic activity could be maintained in most cases, yet the enzymatically active ...
    Detection of Catalase Activity by Polyacrylamide Gel Electrophoresis (PAGE) in Cell Extracts from Pseudomonas aeruginosa
    Authors:  Magdalena Pezzoni, Ramón A. Pizarro and Cristina S. Costa, date: 06/05/2018, view: 183, Q&A: 0
    [Abstract] Bacteria in nature and as pathogens commonly face oxidative stress which causes damage to proteins, lipids and DNA. This damage is produced by the action of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2), singlet oxygen, superoxide anion and hydroxyl radical. ROS are generated by antimicrobials, environmental ...
    In vitro Analysis of Ubiquitin-like Protein Modification in Archaea
    Authors:  Xian Fu, Zachary Adams and Julie A. Maupin-Furlow, date: 05/20/2018, view: 324, Q&A: 0
    [Abstract] The ubiquitin-like (Ubl) protein is widely distributed in Archaea and involved in many cellular pathways. A well-established method to reconstitute archaeal Ubl protein conjugation in vitro is important to better understand the process of archaeal Ubl protein modification. This protocol describes the in vitro reconstitution of ...
    Quantification of Salicylic Acid (SA) and SA-glucosides in Arabidopsis thaliana
    [Abstract] Homeostasis between the cytoplasmic plant hormone salicylic acid (SA) and its’ inactive, vacuolar storage forms, SA-2-O-β-D-glucoside (SAG) and SA-β-D-Glucose Ester (SGE), regulates the fine-tuning of defense responses to biotrophic pathogens in Arabidopsis thaliana. This protocol describes a simplified, optimized procedure to ...