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Past Issues in 2012

Volume: 2 Issue: 20

Biochemistry

Isolate and Sub-fractionate Cell Membranes from Caulobacter crescentus Author: Khatira Anwari, date: 10/20/2012, view: 10661, Q&A: 1

Cell membranes from Caulobacter can be isolated and separated into inner and outer membranes according their characteristic buoyant densities on a sucrose gradient. Fractionation can be used to determine the localisation of uncharacterised proteins and to enrich protein complexes present in either of these membranes for biochemical ...

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Immunology

Schistosoma haematobium Egg Isolation Authors: Chi-Ling Fu and Michael H. Hsieh, date: 10/20/2012, view: 8875, Q&A: 1

Schistosoma haematobiumis the etiologic agent for urogenital schistosomiasis, a major source of morbidity and mortality for more than 112 million people worldwide. Infection with S. haematobium results in a variety of immunopathologic sequelae caused by parasite oviposition within the urinary tract, which drives inflammation, ...

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Microbiology

Minimum Inhibitory Concentration (MIC) Assay for Antifungal Drugs Authors: Jinglin L. Xie, Sheena D. Singh-Babak and Leah E. Cowen, date: 10/20/2012, view: 26011, Q&A: 0

The Minimum Inhibitory Concentration (MIC) Assay is widely used to measure the susceptibility of yeasts to antifungal agents. In serial two-fold dilutions, the lowest concentration of antifungal drug that is sufficient to inhibit fungal growth is the MIC. Typically, 50% inhibitory (MIC₅₀) or 80% inhibitory (MIC₈₀) values are ...

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Differential in vivo Thiol Trapping with N-ethylmaleimide (NEM) and 4-acetamido-4'-maleimidylstilbene-2,2'-disulfonic acid (AMS) Author: Wei-Yun (Winnie) Wholey, date: 10/20/2012, view: 11904, Q&A: 0

This protocol is used to compare the in vivo redox status of Escherichia coli and Vibrio cholerae protein before and after HOCl treatment. For example, I examined whether the EF-Tu protein is reduced or oxidized in the referenced publication. This protocol should work for other proteins and other oxidative stress ...

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Isolation of Inner Membrane Vesicles from Escherichia coli by Using an Affinity Tag Authors: Gang Li and Kevin D. Young, date: 10/20/2012, view: 9386, Q&A: 0

This protocol was developed in a project aimed to identify the inner membrane proteins localizing to cell poles in Escherichia coli (E. coli). By using a known polar protein Tar as a tag, we isolated pole-derived inner membrane vesicles by affinity capture. The specificity of the polar vesicle isolation was confirmed by mass ...

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Promoter Orientation of Prokaryotic Phase-variable Genes by PCR Authors: Stacey L. Bateman and Patrick Seed, date: 10/20/2012, view: 7737, Q&A: 0

One major mechanism of phase variable gene expression in prokaryotes is through inversion of the promoter element for a gene or operon. This protocol describes how to detect the promoter orientation of a phase-variable gene by PCR. This protocol, including primer design, is specific to detection of the promoter orientations of hyxR, a LuxR-like ...

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Intracellular Macrophage Infections with E. coli under Nitrosative Stress Authors: Stacey L. Bateman and Patrick Seed, date: 10/20/2012, view: 11184, Q&A: 0

Escherichia coli (E. coli) produces disseminated infections of the urinary tract, blood, and central nervous system where it encounters professional phagocytes such as macrophages, which utilize reactive nitrogen intermediates (RNI) to arrest bacteria. In vitro, extraintestinal pathogenic E. coli (ExPEC) can ...

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Plant Science

Glycolate Oxidase Activity Assay in Plants Authors: Amita Kaundal, Clemencia M. Rojas and Kirankumar S. Mysore, date: 10/20/2012, view: 9770, Q&A: 0

Glycolate oxidase is located in the peroxisome and is involved in the photorespiratory cycle which recovers some of the carbon loss during photosynthesis. Glycolate oxidase converts glycolate to glyxoylate with the concomitant production of H₂O₂.In this assay, the H₂O₂ generated, in the presence of HRP, ...

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Measurement of NADPH Oxidase Activity in Plants Authors: Amita Kaundal, Clemencia M. Rojas and Kirankumar S. Mysore, date: 10/20/2012, view: 16383, Q&A: 1

NADPH oxidase is a membrane-bound enzyme that generates (O^2-) by transferring electrons from NADPH to molecular oxygen O₂. O^2-is spontaneously dismasted to the more stable form H₂O₂. Both O^2-and H₂O₂ are forms ofreactive oxygen species (ROS), which are involved ...

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