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Past Issues in 2016

Volume: 6 Issue: 1

Cancer Biology

Measurement of 2-methylthio Modifications in Mitochondrial Transfer RNAs by Reverse-transcription Quantitative PCR Authors: Fan-Yan Wei and Kazuhito Tomizawa, date: 01/05/2016, view: 6654, Q&A: 0

2-Methylthio-N⁶-isopentenyladenosine (ms²i⁶A) is an evolutionally conserved posttranscriptional modification found at position 37 of four mammalian mitochondrial tRNAs, mt-tRNA^Ser(UCN), mt-tRNA^Trp, mt-tRNA^Phe and mt-tRNA^Tyr. The ms² modification in ms² ...

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Preparation of Recombinant Galectin-3 for Cancer Studies Authors: Kari Tyler, Sok-Hyong Lee and Erwin G. Van Meir, date: 01/05/2016, view: 7436, Q&A: 0

Galectin-3 is a member of a class of proteins termed Galectins, characterized by their ability to bind glycans containing β-galactose (Cummings and Liu, 2009). Galectin-3 binds preferentially to proteoglycans terminating with N-acetyllactosamine (LacNAc) chains (i.e., tandem repeats of galactose) (Newlaczyl and Yu, 2011). Galectin-3 is ...

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PhagoKinetic Track Assay: Imaging and Analysis of Single Cell Migration Authors: Michiel Fokkelman, Wies van Roosmalen, Vasiliki-Maria Rogkoti, Sylvia E. Le Dévédec, Benjamin Geiger and Bob van de Water, date: 01/05/2016, view: 8004, Q&A: 0

Cell migration is a highly complex and dynamic biological process, essential in several physiological phenomena and pathologies including cancer dissemination and metastasis formation. Thus understanding single cell migration is highly relevant and requires a suitable image-based assay. Depending on the speed of the moving cells, one may require ...

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Immunology

Protocol-In vitro T Cell Proliferation and Treg Suppression Assay with Celltrace Violet Authors: Kristofor K. Ellestad and Colin C. Anderson, date: 01/05/2016, view: 25491, Q&A: 0

Measurement of the incorporation of radionuclides such as ³H-thymidine is a classical immunological technique for assaying T cell proliferation. However, such an approach has drawbacks beyond the inconvenience of working with radioactive materials, such as the inability of bulk radionuclide incorporation measurements to accurately ...

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Neuroscience

Neurite Outgrowth Assay Authors: Angela R. Filous and Jerry Silver, date: 01/05/2016, view: 9155, Q&A: 0

Neurite outgrowth in culture provides an easy way to determine the effects of a particular substrate or exogenous factor on neuron behavior. Dissociated neurons can be plated on a variety of substrates and the length of the longest neurite outgrowth can be compared. Here, we describe how to isolate and dissociate dorsal root ganglion (DRG) ...

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Fluoro-Jade B Staining for Neuronal Cell Death Authors: Nathalie Laflamme, Paul Préfontaine and Serge Rivest, date: 01/05/2016, view: 19012, Q&A: 2

Fluoro-Jade is a fluorescent derivative used for histological staining of degenerating neurons. This technique is simple and sensitive enough to label distal dendrites, axons, axon terminals as well as neuronal bodies. Fluoro-Jade has excitation and emission peak of 480 and 525 nanometer respectively. It can be visualized using a fluorescein/FITC ...

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Isolation and Purification of Murine Microglial Cells for Flow Cytometry Authors: Peter Theriault, Maude Bordeleau and Serge Rivest, date: 01/05/2016, view: 11717, Q&A: 0

The detailed protocol is used to isolate different cell types from murine brain as glial cells, including microglia, using an enzymatic digestion that minimizes cellular mortality. A Percoll gradient (30% to 80%) separation allows a maximal recovery of isolated murine microglial cells prior to flow cytometry analysis.

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Plant Science

Super-resolution Imaging of Live BY2 Cells Using 3D-structured Illumination Microscopy Authors: Karen Bell, Karl Oparka and Kirsten Knox, date: 01/05/2016, view: 7201, Q&A: 0

Light microscopy is the standard tool for studying sub-cellular structures however, owing to the diffractive properties of light, resolution is limited to 200 nm. Super-resolution microscopy methods circumvent this limit, offering greater resolution, particularly when studying fluorescently labeled sub-cellular structures. Super-resolution methods ...

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Measurement of Uptake and Root-to-Shoot Distribution of Sulfate in Arabidopsis Seedlings Authors: Naoko Yoshimoto, Tatsuhiko Kataoka, Akiko Maruyama-Nakashita and Hideki Takahashi, date: 01/05/2016, view: 7439, Q&A: 0

Sulfur is an essential macronutrient required for growth and development of plants. Plants take up sulfate from the soil environment through the function of plasma membrane-bound sulfate transporters expressed at the root surface cell layers. Plants then utilize the incorporated sulfate as the main sulfur source to synthesize sulfur-containing ...

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Saccharification Protocol for Small-scale Lignocellulosic Biomass Samples to Test Processing of Cellulose into Glucose Authors: Rebecca Van Acker, Ruben Vanholme, Kathleen Piens and Wout Boerjan, date: 01/05/2016, view: 9527, Q&A: 0

Second generation biofuels are derived from inedible lignocellulosic biomass of food and non-food crops. Lignocellulosic biomass is mainly composed of cell walls that contain a large proportion of cellulosic and hemicellulosic polysaccharides. An interesting route to generate biofuels and bio-based materials is via enzymatic hydrolysis of cell ...

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Quantification of Low Molecular Weight Thiols in Arabidopsis Authors: Ziqing Miao, Zhen Wang and Cheng-Bin Xiang, date: 01/05/2016, view: 6107, Q&A: 0

Low-molecular-weight (LMW) thiols are a class of highly reactive compounds due to their thiol moiety. They play important roles in the maintenance of cellular redox homeostasis, detoxification, and development. Monobromobimane (mBBr) is weakly fluorescent but selectively reacts with thiols to yield highly fluorescent thioethers (mBSR) products, ...

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