• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

Lipid Extraction from Mouse Feces Authors:  Daniel Kraus, Qin Yang and Barbara B. Kahn, date: 01/05/2015, view: 11955, Q&A: 0
The analysis of feces composition is important for the study of energy metabolism, which comprises various measurements of energy intake, energy expenditure, and energy wasting. The current protocol describes how to measure energy-dense lipids in mouse feces using a modification of the method proposed by Folch et al. (1957).
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Cancer Biology

Three Dimensional Spheroid Co-culture Invasion Assay Authors:  Eldo T Verghese and Thomas A Hughes, date: 01/05/2015, view: 9182, Q&A: 0
The assay was developed to investigate the impact of stromal cells of different types (in our case breast cancer associated fibroblasts stably manipulated to modify expression of genes of interest) on the invasive capacity of epithelial cancer cells (in our case breast cancer cell lines) (Verghese et al., 2013). Typical two dimensional ...
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Analysis of Protein Stability by the Cycloheximide Chase Assay Authors:  Shih-Han Kao, Wen-Lung Wang, Chi-Yuan Chen, Yih-Leong Chang, Yi-Ying Wu, Yi-Ting Wang, Shu-Ping Wang, Alexey I Nesvizhskii, Yu-Ju Chen, Tse-Ming Hong and Pan-Chyr Yang, date: 01/05/2015, view: 47890, Q&A: 1
Comparison of protein stability in eukaryotic cells has been achieved by cycloheximide, which is an inhibitor of protein biosynthesis due to its prevention in translational elongation. It is broadly used in cell biology in terms of determining the half-life of a given protein and has gained much popularity in cancer research. Here we present a ...
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Immunology

Intracellular Cytokine Staining on PBMCs Using CyTOFTM Mass Cytometry Authors:  Dongxia Lin, Sheena Gupta and Holden Maecker, date: 01/05/2015, view: 14500, Q&A: 2
In this protocol, we use a CyTOFTM mass cytometry to collect single-cell data on a large number of cytokines/chemokines as well as cell-surface proteins that characterize T cells and other immune cells. The current selected mass window in AW 103-203 includes the lanthanides used for most antibody labeling, along with iridium and rhodium ...
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Microbiology

Quantification of Extracellular Ammonium Concentrations and Transporter Activity in Yeast Using AmTrac Fluorescent Sensors Authors:  Cindy Ast, Wolf B. Frommer, Guido Grossmann and Roberto De Michele, date: 01/05/2015, view: 7592, Q&A: 0
AmTracs are the first example of “activity sensors”, since they report the activity of ammonium transporters by means of fluorescence readout in vivo (De Michele et al., 2013). AmTracs are based on a single fluorescent protein, a circularly permuted GFP (cpGFP), inserted into the cytosolic loop connecting the two ...
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Construction of FWPV Chimaeric MVA Authors:  Karen R. Buttigieg and Michael A. Skinner, date: 01/05/2015, view: 5692, Q&A: 1
Construction of chimaeric MVA is a useful tool with which to study gene function of related viruses. The protocol given here describes MVA chimaeras containing genes from Fowlpox virus (FWPV), although this can be applied to DNA derived from other organisms. There are a number of steps required to make the chimaeric MVA: 1) Purification of viral ...
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Neuroscience

Manganese Cytotoxicity Assay on Hippocampal Neuronal Cell Culture Authors:  Alexia Daoust, Yasmina Saoudi, Jacques Brocard, Nora Collomb, Cécile Batandier, Mariano Bisbal, Murielle Salomé, Annie Andrieux, Sylvain Bohic and Emmanuel L. Barbier, date: 01/05/2015, view: 6067, Q&A: 0
Compared to an in vivo experiment, neuronal cell cultures are immediately accessible to observation and manipulation. In this protocol, we describe a technique to evaluate the cytotoxicity of a metal, manganese (Mn2+), on hippocampal neuronal cell cultures. Interestingly, this protocol is easily adaptable to any type of primary ...
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In vitro Migration Assays for Neural Stem Cells, Intermediate Neurogenic Progenitors and Immature Neurons Authors:  Julia Ladewig, Philipp Koch and Oliver Brüstle, date: 01/05/2015, view: 11825, Q&A: 0
In the vertebrate central nervous system (CNS), different neural precursor populations such as neural stem cells (NSCs), intermediate neurogenic progenitors (INPs) and immature neurons have to migrate from their places of birth to their location of function. Coordinated migration is mediated by direct cell-cell interactions and by extracellular ...
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Plant Science

Collection of Root Exudate from Duckweed Authors:  Yufang Lu, Li Sun and Weiming Shi, date: 01/05/2015, view: 9675, Q&A: 0
Root exudates play an important role in rhizosphere interactions between plants and microorganisms. However, collection of chemicals from plant root system is difficult due to their low concentrations and high level of contaminants in growth media. The continuous collection method has been described in several terrestrial plants over the past 30 ...
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Imaging and Quantitative Analysis of Size and Distribution of Spherical Bodies, e.g. Embryonic Oil Bodies Authors:  Martine Miquel, Ghassen Trigui, Alain Trubuil and Bertrand Dubreucq, date: 01/05/2015, view: 5960, Q&A: 0
Oil bodies (OBs) are seed-specific lipid storage organelles that allow the accumulation of neutral lipids that sustain plantlet development after the onset of germination. Using fluorescent dyes and confocal microscopy, we monitored the dynamics of OBs in living Arabidopsis (Arabidopsis thaliana) embryos at different stages of development ...
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