Volume: 8 Issue: 15 —BIO-PROTOCOL

Biochemistry

Isothermal Titration Calorimetry: A Biophysical Method to Characterize the Interaction between Label-free Biomolecules in Solution Author: Andrea Saponaro, date: 08/05/2018, view: 15380, Q&A: 0

This protocol can be applied to analyze the direct interaction between a soluble protein and a target ligand molecule using Isothermal Titration Calorimetry (ITC, Malvern). ITC allows the biophysical characterization of binding between label-free, non-immobilized and in-solution biomolecules by providing the stoichiometry of the interaction, the ...

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Inositol Phosphates Purification Using Titanium Dioxide Beads Authors: Miranda SC Wilson and Adolfo Saiardi, date: 08/05/2018, view: 3082, Q&A: 0

Inositol phosphates (IPs) comprise a family of ubiquitous eukaryotic signaling molecules. They have been linked to the regulation of a pleiotropy of important cellular activities, but low abundance and detection difficulties have hampered our understanding. Here we present a method to purify and enrich IPs or other phosphate-rich metabolites from ...

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Cancer Biology

Hypoxia Reporter Element Assay Authors: Daelynn R Buelow and Sharyn D. D Baker, date: 08/05/2018, view: 3601, Q&A: 0

Hypoxia is a condition in which there is a decrease in oxygen supply to the cellular environment. Changes to cellular oxygen levels can lead to transcriptional changes of oxygen-regulated genes. Reporter assays are used to study gene expression alteration and modifications in response to environmental changes. Dual-reporter assays allow the ...

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Developmental Biology

Isolation of LYVE-1⁺ Endothelial Cells from Mouse Embryos Author: Patrick Crosswhite, date: 08/05/2018, view: 3233, Q&A: 0

Lymphatic vessel endothelial hyaluronan receptor 1, or LYVE-1, is a type 1 integral membrane glycoprotein expressed by lymphatic endothelial cells (LECs). LYVE-1 is commonly used as a biological marker to visually distinguish developing lymphatic vessels from blood endothelial cells (arteries or veins). As our understanding of lymphatic biology is ...

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Microbiology

Long-term in vitro Culture of Cryptosporidium parvum Authors: Nigel Yarlett and Mary Morada, date: 08/05/2018, view: 4249, Q&A: 0

Continuous in vitro growth of Cryptosporidium parvum has proved difficult and conventional in vitro culture techniques result in short-term (2-5 days) growth of the parasite resulting in thin-walled oocysts that fail to propagate using in vitro cultures, and do not produce an active infection using ...

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Detection of Apoptosis-like Cell Death in Ustilago maydis by Annexin V-FITC Staining Authors: Dibya Mukherjee, Aroni Mitra and Anupama Ghosh, date: 08/05/2018, view: 3435, Q&A: 0

Programmed cell death (PCD) guides the transition between key developmental stages in many organisms. PCD also remains an important fate for many organisms upon exposure to different stress conditions. Therefore, an insight into the progression of PCD during the execution of a biological phenomenon can yield significant details of the underlying ...

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HCV Reporter System (Viral Infection-Activated Split-Intein-Mediated Reporter System) for Testing Virus Cell-to-cell Transmission ex-vivo Authors: Fanfan Zhao, Ting Zhao, Libin Deng, Dawei Lv, Xiaolong Zhang, Xiaoyu Pan, Jun Xu and Gang Long, date: 08/05/2018, view: 3288, Q&A: 0

Hepatitis C virus (HCV) spread involves two distinct entry pathways: cell-free transmission and cell-to-cell transmission. Cell-to-cell transmission is not only an efficient way for viruses to spread but also an effective method for escaping neutralizing antibodies. We adapted the viral infection-activated split-intein-mediated reporter system ...

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Structural Analysis of Bordetella pertussis Biofilms by Confocal Laser Scanning Microscopy Authors: Natalia Cattelan, Osvaldo Miguel Yantorno and Rajendar Deora, date: 08/05/2018, view: 3536, Q&A: 0

Biofilms are sessile communities of microbial cells embedded in a self-produced or host-derived exopolymeric matrix. Biofilms can both be beneficial or detrimental depending on the surface. Compared to their planktonic counterparts, biofilm cells display enhanced resistance to killing by environmental threats, chemicals, antimicrobials and host ...

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Direct Visualization of the Multicopy Chromosomes in Cyanobacterium Synechococcus elongatus PCC 7942 Authors: Ryudo Ohbayashi, Hirofumi Yoshikawa and Satoru Watanabe, date: 08/05/2018, view: 3942, Q&A: 0

Cyanobacteria are prokaryotic organisms that carry out oxygenic photosynthesis. The fresh water cyanobacterium Synechococcus elongatus PCC 7942 is a model organism for the study of photosynthesis and gene regulation, and for biotechnological applications. Besides several freshwater cyanobacteria, S. elongatus 7942 also contains ...

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Fusarium graminearum Inoculation on Wheat Head Authors: Chanjing Feng, Huiquan Liu and Zhe Tang, date: 08/05/2018, view: 4638, Q&A: 0

Fusarium graminearum, the major causal agent of Fusarium head blight (FHB), causes serious wheat yield losses and a threat to human and animal health. The main efforts to combat the disease are the research of pathogenesis mechanisms and breeding for disease resistance plants. The efficiency of these actions could be evaluated by ...

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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: 5356, Q&A: 0

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 ...

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Neuroscience

Quantitative Electron Microscopic Assay Using Random Sampling from Single Sections to Test Plastic Synaptic Changes in Hippocampus Authors: G. Mark Marcello, Lilla E. Szabó, Peter Sotonyi and Bence Rácz, date: 08/05/2018, view: 3202, Q&A: 0

Studies over several decades on the organization of the CA1 hippocampus–a particularly favorable model for learning, memory and certain forms of cognition–have shown that the synaptic network in this brain region is plastic (Fortin et al., 2012). Recent evidence suggests that a number of environmental and endogenous stimuli may have a ...

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Plant Science

Arabidopsis-Green Peach Aphid Interaction: Rearing the Insect, No-choice and Fecundity Assays, and Electrical Penetration Graph Technique to Study Insect Feeding Behavior Authors: Vamsi Nalam, Joe Louis, Monika Patel and Jyoti Shah, date: 08/05/2018, view: 5313, Q&A: 1

Aphids constitute a large group of Hemipterans that use their slender stylets to tap into the sieve elements of plants from which they consume copious amounts of phloem sap, thus depriving the plant of photoassimilates. Some aphids also transmit viral diseases of plants. Myzus persicae Sülzer, commonly known as the green peach aphid ...

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Measuring Secretion of Capsidiol in Leaf Tissues of Nicotiana benthamiana Authors: Teruhiko Kuroyanagi, Maurizio Camagna and Daigo Takemoto, date: 08/05/2018, view: 3065, Q&A: 0

Plant species produce a wide variety of antimicrobial metabolites to protect themselves against potential pathogens in natural environments. Phytoalexins are low molecular weight compounds produced by plants in response to attempted attacks of pathogens. Accumulation of phytoalexins in attacked plant tissues can inhibit the growth of penetrating ...

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Tethered Chromosome Conformation Capture Sequencing in Triticeae: A Valuable Tool for Genome Assembly Authors: Axel Himmelbach, Ines Walde, Martin Mascher and Nils Stein, date: 08/05/2018, view: 4787, Q&A: 0

Chromosome conformation capture sequencing (Hi-C) is a powerful method to comprehensively interrogate the three-dimensional positioning of chromatin in the nucleus. The development of Hi-C can be traced back to successive increases in the resolution and throughput of chromosome conformation capture (3C) (Dekker et al., 2002). The basic ...

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Past Issues in 2018

Volume: 8 Issue: 15

Biochemistry

Cancer Biology

Developmental Biology

Microbiology

Neuroscience

Plant Science