Biochemistry

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    Protocols in Current Issue
    Photoaffinity Labeling of Respiratory Complex I in Bovine Heart Submitochondrial Particles by Photoreactive [125I] amilorides
    Authors:  Masatoshi Murai and Hideto Miyoshi, date: 09/05/2019, view: 170, Q&A: 0
    [Abstract] The architecture of quinone/inhibitor-access channel in proton-translocating NADH-quinone oxidoreductase (respiratory complex I) was modeled by X-ray crystallography and cryo-EM, however, it remains debatable whether the channel model reflects the physiologically relevant state present throughout the catalytic cycle. Using photoreactive [125 ...
    Quantification of Prostaglandin E2 Concentration in Interstitial Fluid from the Hypothalamic Region of Free-moving Mice
    Authors:  Yoshinobu Nakamura, Takatoshi Sakaguchi, Ikumi Tamai and Takeo Nakanishi, date: 08/05/2019, view: 352, Q&A: 0
    [Abstract] Prostaglandin E2 (PGE2) is a well-established chemical mediator for the generation of the fever at the hypothalamus of the brain. PGE2 mediates fever generation via PGE receptor 3 (i.e., EP3) on neurons in the preoptic area. The role of PGE2 has been analyzed by measuring PGE2 ...
    Oxygen Consumption Measurements in Caenorhabditis elegans Using the Seahorse XF24
    Authors:  Suraiya Haroon and Marc Vermulst, date: 07/05/2019, view: 556, Q&A: 0
    [Abstract] Mitochondria generate 90% of the energy required to sustain life. As a result, loss of mitochondrial function compromises almost every facet of human physiology. Accordingly, most mitochondrial diseases tend to present themselves as complex, multi-systemic disorders that can be difficult to diagnose. Depending on the severity of the mitochondrial ...
    Simple Method to Determine Protein Redox State in Arabidopsis thaliana
    Authors:  Keisuke Yoshida and Toru Hisabori, date: 06/05/2019, view: 1106, Q&A: 0
    [Abstract] Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...
    Hypochlorous Acid Staining with R19-S in the Drosophila Intestine upon Ingestion of Opportunistic Bacteria
    Authors:  Salma Hachfi, Olivia Benguettat and Armel Gallet, date: 05/20/2019, view: 812, Q&A: 0
    [Abstract] The intestine is endowed with an innate immune system that is required to fight any exogenous bacteria that are swallowed along with the food. The first line of defense that is mounted by the gut epithelium is the release of immune Reactive Oxygen Species (ROS), such as hypochlorous acid (HOCl), into the lumen. HOCl is produced within 1.5 h of ...
    An Adjustable Protocol to Analyze Chemical Profiles of Non-sterile Rhizosphere Soil
    Authors:  Alex Williams, Jurriaan Ton and Pierre Pétriacq, date: 05/20/2019, view: 1276, Q&A: 0
    [Abstract] The analysis of chemical diversity in non-sterile rhizosphere soil has been a pressing methodological challenge for years. Rhizosphere-enriched chemicals (i.e., rhizochemicals) include root exudation chemicals, (microbial) breakdown products thereof, and de novo produced metabolites by rhizosphere-inhabiting microbes, all of ...
    An Improved Bioassay to Study Arabidopsis Induced Systemic Resistance (ISR) Against Bacterial Pathogens and Insect Pests
    Authors:  Nicolás M. Cecchini, Yi Song, Suruchi Roychoudhry, Jean T. Greenberg and Cara H. Haney, date: 05/20/2019, view: 1553, Q&A: 0
    [Abstract] The plant immune system is essential for plants to perceive and defend against bacterial, fungal and insect pests and pathogens. Induced systemic resistance (ISR) is a systemic immune response that occurs upon root colonization by beneficial microbes. A well-studied model for ISR is the association of specific beneficial strains of Pseudomonas ...
    Analysis of Indole-3-acetic Acid (IAA) Production in Klebsiella by LC-MS/MS and the Salkowski Method
    Authors:  Shraddha Gang, Sheetal Sharma, Meenu Saraf, Martin Buck and Jörg Schumacher, date: 05/05/2019, view: 1564, Q&A: 0
    [Abstract] Many rhizobacteria isolated from plant rhizosphere produce various phytohormones in the form of secondary metabolites, the most common of which is Indole-3-acetic acid (IAA). Here, we detail analytical protocols of IAA detection and quantification, in vitro and in situ, as recently applied to Klebsiella SGM 81, a ...
    Optimized Oxidative Stress Protocols for Low-microliter Volumes of Mammalian Plasma
    Author:  Evan A. Langille, date: 05/05/2019, view: 813, Q&A: 0
    [Abstract] Small blood volumes commonly obtained from small mammals during field studies are only sufficient for a single biochemical assay. In this study, we used blood collected from a population of wild eastern chipmunks (Tamias striatus) and developed modified methods to improve analytical selectivity and sensitivity required for measuring ...
    Quantification of the Humidity Effect on HR by Ion Leakage Assay
    Authors:  Musoki Mwimba and Xinnian Dong, date: 04/05/2019, view: 905, Q&A: 0
    [Abstract] We describe a protocol to measure the contribution of humidity on cell death during the effector-triggered immunity (ETI), the plant immune response triggered by the recognition of pathogen effectors by plant resistance genes. This protocol quantifies tissue cell death by measuring ion leakage due to loss of membrane integrity during the ...



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