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

Current Issue in 2018

Volume: 8, Issue: 13


Assessment of Uptake and Biodistribution of Radiolabeled Cholesterol in Mice Using Gavaged Recombinant Triglyceride-rich Lipoprotein Particles (rTRL) Authors:  Anna Worthmann, Clara John and Joerg Heeren, date: 07/05/2018, view: 170, Q&A: 0
The here described method can be used to estimate the uptake of orally provided cholesterol in mice. Briefly, mice are gavaged with radiolabeled cholesterol and 4 h later, organ distribution of the radiolabel is determined by liquid scintillation counting. The method has been applied successfully to determine dietary cholesterol handling of mice ...
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Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging Author:  Andrew D. D Doyle, date: 07/05/2018, view: 252, Q&A: 0
Rat tail collagen solutions have been used as polymerizable in vitro three-dimensional (3D) extracellular matrix (ECM) gels for single and collective cell migration assays as well as spheroid formation. These 3D hydrogels are a relatively inexpensive, simple to use model system that can mimic the in vivo physical characteristics ...
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Cancer Biology

An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist Authors:  Khalidah Ibraheem, Christopher J. Dunnill, Myria Ioannou, Albashir Mohamed, Balid Albarbar and Nikolaos T. Georgopoulos, date: 07/05/2018, view: 288, Q&A: 0
One fundamental property of the TNR receptor (TNFR) family relates to how ‘signal quality’ (the extent of receptor ligation or cross-linking) influences the outcome of receptor ligation, for instance the induction of death in tumour cells. It is unequivocal that membrane-presented ligand (delivered to target cells via cell-surface presentation by ...
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Cell Biology

FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans Authors:  Ranjan Devkota and Marc Pilon, date: 07/05/2018, view: 180, Q&A: 0
FRAP (Fluorescence Recovery After Photobleaching) is probably the most direct method to investigate the dynamics of molecules in living cells. Here, we describe FRAP to quantify membrane fluidity in C. elegans. Using FRAP, we have shown that cold, glucose and exogenous saturated fatty acids can decrease the fluidity of cellular membranes ...
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Developmental Biology

Preserve Cultured Cell Cytonemes through a Modified Electron Microscopy Fixation Authors:  Eric T. Hall and Stacey K. Ogden, date: 07/05/2018, view: 150, Q&A: 0
Immunocytochemistry of cultured cells is a common and effective technique for determining compositions and localizations of proteins within cellular structures. However, traditional cultured cell fixation and staining protocols are not effective in preserving cultured cell cytonemes, long specialized filopodia that are dedicated to morphogen ...
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Mouse Mammary Gland Whole Mount Preparation and Analysis Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 07/05/2018, view: 145, Q&A: 0
The mammary gland undergoes extensive remodeling during pregnancy and is also subject to neoplastic processes both of which result in histological changes of the gland epithelial structure. Since the mammary tree is a complex three-dimensional structure a method is needed that provides an overview of the entire gland. Whole mounts provide this ...
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Sleeping Beauty Transposon-based System for Rapid Generation of HBV-replicating Stable Cell Lines Authors:  Jin-Wei Zheng, Jia-Li Cao and Quan Yuan , date: 07/05/2018, view: 142, Q&A: 0
The stable HBV-transfected cell lines, which based on stable integration of replication-competent HBV genome into hepatic cells, are widely used in basic research and antiviral drug evaluation against HBV. However, previous reported strategies to generate HBV-replicating cell lines, which primarily rely on random integration of exogenous DNA by ...
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Bacterial Microcolonies in Gel Beads for High-throughput Screening Author:  Yolanda Schaerli, date: 07/05/2018, view: 758, Q&A: 0
High-throughput screening of a DNA library expressed in a bacterial population for identifying potentially rare members displaying a property of interest is a crucial step for success in many experiments such as directed evolution of proteins and synthetic circuits and deep mutational scanning to identify gain- or loss-of-function mutants.
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Induction of Natural Competence in Genetically-modified Lactococcus lactis Authors:  Joyce Mulder, Michiel Wels, Oscar P. Kuipers, Michiel Kleerebezem and Peter A. Bron, date: 07/05/2018, view: 98, Q&A: 0
Natural competence can be activated in Lactoccocus lactis subsp lactis and cremoris upon overexpression of ComX, a master regulator of bacterial competence. Herein, we demonstrate a method to activate bacterial competence by regulating the expression of the comX gene by using a nisin-inducible promoter in an L. ...
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Brain Tissue Culture of Per2::Luciferase Transgenic Mice for ex vivo Bioluminescence Authors:  Nora L. Salaberry and Jorge Mendoza, date: 07/05/2018, view: 157, Q&A: 0
In circadian research, it is essential to be able to track a biological rhythm for several days with the minimum perturbation for the organisms or tissues. The use of transgenic mice lines, in which the luciferase reporter is coupled to a molecular clock protein (here PERIOD2), gives us the opportunity to follow the circadian activity in different ...
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Preparation of Cerebellum Granule Neurons from Mouse or Rat Pups and Evaluation of Clostridial Neurotoxin Activity and Their Inhibitors by Western Blot and Immunohistochemistry Authors:  Domenico Azarnia Tehran and Marco Pirazzini, date: 07/05/2018, view: 175, Q&A: 0
Cerebellar Granule Neurons (CGN) from post-natal rodents have been widely used as a model to study neuronal development, physiology and pathology. CGN cultured in vitro maintain the same features displayed in vivo by mature cerebellar granule cells, including the development of a dense neuritic network, neuronal activity, ...
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Plant Science

Separation of Thylakoid Protein Complexes with Two-dimensional Native-PAGE Authors:  Marjaana Rantala, Virpi Paakkarinen and Eva-Mari Aro, date: 07/05/2018, view: 252, Q&A: 0
The hierarchical composition and interactions of the labile thylakoid protein complexes can be assessed by sequential 2D-native gel-electrophoresis system. Mild non-ionic detergent digitonin is used to solubilize labile protein super-and megacomplexes, which are then separated with first-dimension blue native polyacrylamide gel electrophoresis ...
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An Optimized CTAB Method for Genomic DNA Extraction from Freshly-picked Pinnae of Fern, Adiantum capillus-veneris L. Authors:  Yi Shu, Jin Wan-Ting, Yuan Ya-Ning and Fang Yu-Han, date: 07/05/2018, view: 159, Q&A: 0
As the sister clade of seed plants, ferns are significant materials for plant phylogeny research. However, the genomic DNA extraction protocol for fern samples like modified CTAB method still lacks robustness. Here, we found that the amount and condition of the pinnae samples are critical for gDNA extraction in fern, Adiantum capillus-veneris ...
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Isolation of Intact Vacuoles from Petunia Petals and Extraction of Sequestered Glycosylated Phenylpropanoid Compounds Authors:  Oded Skaliter , Jasmin Ravid, Alon Cna'ani, Gony Dvir, Rafael Knafo and Alexander Vainstein, date: 07/05/2018, view: 133, Q&A: 0
Plant vacuoles are the largest compartment in plant cells, occupying more than 80% of the cell volume. A variety of proteins, sugars, pigments and other metabolites are stored in these organelles (Paris et al., 1996; Olbrich et al., 2007). Flowers produce a variety of specialized metabolites, some of which are unique to this ...
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A Modified Approach for Axenic Cultivation of Spores of Fern Adiantum capillus-veneris L. with High Germination Rate Authors:  Yuan Ya-Ning, Yi Shu, Jin Wan-Ting and Fang Yu-Han, date: 07/05/2018, view: 142, Q&A: 0
Spores are the primary way of spread and reproduction for ferns, a clade of seed-free vascular plants. However, no detailed protocol for ferns spore cultivation has been reported yet. Here we provide a modified approach for axenic cultivation of fern Adiantum capillus-veneris L., based on Cao’s and Li’s method (Cao, et al., 2010; ...
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Quantification of Starch in Guard Cells of Arabidopsis thaliana Authors:  Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 212, Q&A: 0
In this protocol, we describe how to quantify starch in guard cells of Arabidopsis thaliana using the fluorophore propidium iodide and confocal laser scanning microscopy. This simple method enables monitoring, with unprecedented resolution, the dynamics of starch in guard cells.
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