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Past Issues in 2016

Volume: 6 Issue: 6

Cancer Biology

In vivo Bioluminescence Imaging of Luciferase-labeled Cancer Cells Authors: Marc Carceles-Cordon, Irene Rodriguez-Fernandez, Veronica Rodriguez-Bravo, Carlos Cordon-Cardo and Josep Domingo-Domenech, date: 03/20/2016, view: 25379, Q&A: 0

Over the past decade, in vivo bioluminescent imaging has emerged as a non-invasive and sensitive tool for studying ongoing biological processes within living organisms (Contag et al., 1997; Contag et al., 1998). Based on the detection and quantitation of the photons produced by the oxidation of luciferin by luciferase ...

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Immunology

In vitro Assessment of Immunological Synapse Formation by Flow Cytometry Authors: Bo-Ra Na and Chang-Duk Jun, date: 03/20/2016, view: 12258, Q&A: 2

In adaptive immune system, formation of immunological synapse between T cells and antigen presenting cells (dendritic cells, B cells, and macrophages) or target cells (tumor cells and viral-infected cells) is critical for the execution of T cell immune responses via cytokine secretion or direct killing activity. Here, we describe the practical ...

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In vitro Differentiation of Murine Innate Lymphoid Cells from Common Lymphoid Progenitor Cells Authors: Corey Seehus and Jonathan Kaye, date: 03/20/2016, view: 6886, Q&A: 0

Subtypes of innate lymphoid cells (ILC), defined based on their cytokine secretion profiles and transcription factor expression, are important for host protection from pathogens and maintaining tissue homeostasis. ILCs develop from common lymphoid progenitors (CLP) in the bone marrow. Using the methods described here, we have previously shown that ...

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Microbiology

In vitro Deneddylation Assay Authors: Anna M. Köhler, Cindy Meister and Gerhard H. Braus, date: 03/20/2016, view: 6635, Q&A: 0

Nedd8 is a small ubiquitin-like protein (9 kDa) covalently attached to a conserved lysine residue of a cullin protein which is part of cullin-RING ligases (CRLs). CRLs are major E3 ligases important for protein ubiquitination in the ubiquitin-proteasome pathway (UPP). The activity of CRLs is regulated by cycles of neddylation (CulA-N8, ~98 kDa) ...

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Detection of Protein Oxidative Activity Using Reduced RNase A Authors: Magdalena Grzeszczuk, Katarzyna Bocian-Ostrzycka, Anna Lasica and E. Katarzyna Jagusztyn-Krynicka, date: 03/20/2016, view: 4958, Q&A: 0

This assay allows to determine whether proteins possess oxidative activity-the ability to introduce disulfide bond in vitro. The substrate for potential oxidases is a ribonuclease A which, for its activity, needs 4 properly formed disulfide bonds (Raines, 1998).
RNase A activity can be detected by:

Monitoring the digestion of ...

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In vitro Fluorescent Matrix Degradation Assay for Entamoeba histolytica Authors: Merlyn Emmanuel and Sunando Datta, date: 03/20/2016, view: 6343, Q&A: 0

Fluorescent matrix degradation assay is a popular and widely used assay in the field of invadopodium biology (Artym et al., 2009). Matrix remodeling and degradation can be observed under both physiological and pathological conditions. Cancer cells extensively remodel and degrade the underlying matrix by employing actin-rich protrusive ...

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Neuroscience

Dissection and Staining of Mouse Brain Ventricular Wall for the Analysis of Ependymal Cell Cilia Organization Authors: Paul Labedan, Cédric Matthews, Laurent Kodjabachian, Harold Cremer, Fadel Tissir and Camille Boutin, date: 03/20/2016, view: 10044, Q&A: 0

In the developing and mature central nervous system (CNS) the ventricular lumen is lined by the neuroepithelium and ependymal, respectively. These ventricular epithelia perform important functions related to the development, morphogenesis and physiology of the brain. In the mature CNS, ependyma constitutes a barrier between brain parenchyma and ...

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Mouse Auditory Brainstem Response Testing Authors: Omar Akil, A. E. Oursler, Kevin Fan and Lawrence R. Lustig, date: 03/20/2016, view: 12098, Q&A: 0

The auditory brainstem response (ABR) test provides information about the inner ear (cochlea) and the central pathways for hearing. The ABR reflects the electrical responses of both the cochlear ganglion neurons and the nuclei of the central auditory pathway to sound stimulation (Zhou et al., 2006; Burkard et al., 2007). The ABR ...

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Plant Science

In vitro Microtubule Binding Assay and Dissociation Constant Estimation Authors: Christopher Kesten, René Schneider and Staffan Persson, date: 03/20/2016, view: 10664, Q&A: 0

Microtubules (MTs) support an astonishing set of versatile cellular functions ranging from cell division, vesicle transport, and cell and tissue morphogenesis in various organisms. This versatility is in large mediated by MT-associated proteins (MAPs). The neuronal MAP Tau, for example, is stabilizing MTs in axons of the vertebrate nervous system ...

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Ustilago maydis Virulence Assays in Maize Authors: Amey Redkar and Gunther Doehlemann, date: 03/20/2016, view: 8150, Q&A: 0

The basidiomycetous smut fungus Ustilago maydis (U. maydis) infects all aerial parts of its host plant maize (Zea mays L.). Infection symptoms are seen in the form of prominent tumors on all aerial parts of maize, after the establishment of a biotrophic interaction with the host usually around 5-6 days post infection ...

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EdU Based DNA Synthesis and Cell Proliferation Assay in Maize Infected by the Smut Fungus Ustilago maydis Authors: Amey Redkar and Gunther Doehlemann, date: 03/20/2016, view: 6609, Q&A: 0

The basidiomycetous smut fungus Ustilago maydis (U. maydis) infects all aerial parts of its host plant maize (Zea mays L.). Infection is seen in the form of prominent tumorous symptoms after the establishment of a biotrophic interaction with the host, usually around 5-6 days after infection. The fungus colonizes the ...

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Extraction and Measurement of Strigolactones in Sorghum Roots Authors: Kaori Yoneyama, Xiaonan Xie, Takahito Nomura and Koichi Yoneyama, date: 03/20/2016, view: 7581, Q&A: 0

Strigolactones (SLs) are carotenoid-derived signaling chemicals containing two lactone moieties in their structures and induce seed germination of root parasitic plants, Striga and Orobanche spp. In the rhizosphere, SLs are essential host recognition signals not only for root parasitic plants but also for arbuscular mycorrhizal ...

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Assay of Arabinofuranosidase Activity in Maize Roots Authors: Liudmila V. Kozlova, Polina V. Mikshina and Tatyana A. Gorshkova, date: 03/20/2016, view: 6600, Q&A: 0

Root is a perfect model for studying the mechanisms of plant cell growth. Along the root length, several zones where cells are at different stages of development can be visualized (Figure 1). The dissection of the root on these zones allows the investigation of biochemical and genetic aspects of different growth steps. Maize primary root is much ...

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Detection of Nitric Oxide and Determination of Nitrite Concentrations in Arabidopsis thaliana and Azospirilum brasilense Authors: Noelia Foresi, Natalia Correa-Aragunde, Melina Amenta, Andrés Arruebarrena, Cecilia Creus and Lorenzo Lamattina, date: 03/20/2016, view: 14932, Q&A: 0

There is now general agreement that nitric oxide (NO) is an important and almost ubiquitous signal in plants. Nevertheless, there are still many controversial observations and differing opinions on the importance and functions of NO in plants. Partly, this may be due to the difficulties in detecting and quantifying NO. Here, we summarize protocols ...

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Sample Preparation for X-ray Micro-computed Tomography of Woody Plant Material and Associated Xylem Visualisation Techniques Author: Nicholas J. B. Brereton, date: 03/20/2016, view: 6921, Q&A: 0

Variation in the tissue structure of short rotation coppice (SRC) willow is a principle factor driving differences in lignocellulosic sugar yield yet much of the physiology and development of this tissue is unknown. Traditional sectioning can be both difficult and destructive in woody tissue; however, technology such as three dimensional X-ray ...

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