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    Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging
    Author:  Andrew D. D Doyle, date: 07/05/2018, view: 208, 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 ...
    Quantification of Starch in Guard Cells of Arabidopsis thaliana
    Authors:  Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 178, 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, ...
    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: 141, 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 ...
    An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist
    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 ...
    Mouse Mammary Gland Whole Mount Preparation and Analysis
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 07/05/2018, view: 114, 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 ...
    Brain Tissue Culture of Per2::Luciferase Transgenic Mice for ex vivo Bioluminescence
    Authors:  Nora L. Salaberry and Jorge Mendoza, date: 07/05/2018, view: 125, 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 ...
    FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans
    Authors:  Ranjan Devkota and Marc Pilon, date: 07/05/2018, view: 138, 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 ...
    Preserve Cultured Cell Cytonemes through a Modified Electron Microscopy Fixation
    Authors:  Eric T. Hall and Stacey K. Ogden, date: 07/05/2018, view: 128, 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 ...
    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: 108, 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. ...
    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: 117, 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 ...
    Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging
    Author:  Andrew D. D Doyle, date: 07/05/2018, view: 208, Q&A: 0
    [Abstract] 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 ...
    Quantification of Starch in Guard Cells of Arabidopsis thaliana
    Authors:  Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 178, Q&A: 0
    [Abstract] 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.
    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: 141, Q&A: 0
    [Abstract] 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, ...
    An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist
    [Abstract] 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 ...
    Mouse Mammary Gland Whole Mount Preparation and Analysis
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 07/05/2018, view: 114, Q&A: 0
    [Abstract] 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 ...
    Brain Tissue Culture of Per2::Luciferase Transgenic Mice for ex vivo Bioluminescence
    Authors:  Nora L. Salaberry and Jorge Mendoza, date: 07/05/2018, view: 125, Q&A: 0
    [Abstract] 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 ...
    FRAP: A Powerful Method to Evaluate Membrane Fluidity in Caenorhabditis elegans
    Authors:  Ranjan Devkota and Marc Pilon, date: 07/05/2018, view: 138, Q&A: 0
    [Abstract] 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 ...
    Preserve Cultured Cell Cytonemes through a Modified Electron Microscopy Fixation
    Authors:  Eric T. Hall and Stacey K. Ogden, date: 07/05/2018, view: 128, Q&A: 0
    [Abstract] 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 ...
    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: 108, Q&A: 0
    [Abstract] 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 ...
    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: 117, Q&A: 0
    [Abstract] 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 ...