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  1. No Access

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    Site-specific Integration of Bacterial Artificial Chromosomes into Human Cells

    Gene therapy of inherited diseases requires long-term maintenance of the corrective transgene. Stable integration of the introduced DNA molecule into one of the host cell chromosomes is the simplest strategy f...

    Bradley McColl, Sara Howden, Jim Vadolas in Bacterial Artificial Chromosomes (2015)

  2. No Access

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    Mitophagy and Mitochondrial Balance

    Mitochondria are highly dynamic organelles, with a morphology ranging from small roundish elements to large interconnected networks. This fine architecture has a significant impact on mitochondrial homeostasis...

    Simone Patergnani, Paolo Pinton Ph.D. in Mitochondrial Regulation (2015)

  3. No Access

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    Hemodynamic Flow Visualization of Early Embryonic Great Vessels Using μPIV

    Microparticle image velocimetry (μPIV) is an evolving quantitative methodology to closely and accurately monitor the cardiac flow dynamics and mechanotransduction during vascular morphogenesis. While PIV techn...

    Selda Goktas, Chia-Yuan Chen, William J. Kowalski in Tissue Morphogenesis (2015)

  4. No Access

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    PCR Based Determination of Mitochondrial DNA Copy Number in Multiple Species

    Mitochondrial DNA (mtDNA) copy number is a critical component of overall mitochondrial health. In this chapter, we describe methods for isolation of both mtDNA and nuclear DNA (nucDNA) and measurement of their...

    John P. Rooney, Ian T. Ryde, Laurie H. Sanders in Mitochondrial Regulation (2015)

  5. No Access

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    A Biochemical Approach to Identify Direct MicroRNA Targets

    We have recently developed a biochemical approach to isolate miRNA-bound mRNAs and have used this method to identify the genome-wide mRNAs regulated by the tumor suppressor miRNA miR-34a. This method involves ...

    Murugan Subramanian, Xiao Ling Li, Toshifumi Hara in Regulatory Non-Coding RNAs (2015)

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    Imaging Callose at Plasmodesmata Using Aniline Blue: Quantitative Confocal Microscopy

    Callose (β-1,3-glucan) is both structural and functional component of plasmodesmata (Pd). The turnover of callose at Pd controls the cell-to-cell diffusion rate of molecules through Pd. An accurate assessment ...

    Raul Zavaliev, Bernard L. Epel in Plasmodesmata (2015)

  7. No Access

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    In Vivo Techniques to Investigate the Internalization Profile of Opioid Receptors

    G-protein-coupled receptors (GPCRs) regulate a remarkable diversity of biological functions, and are thus often targeted for drug therapies. Receptor internalization is commonly observed following agonist bind...

    Amynah A. Pradhan, Vivianne L. Tawfik, Alycia F. Tipton in Opioid Receptors (2015)

  8. No Access

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    Determination of the Dissociation Constant of the NFκB p50/p65 Heterodimer in Living Cells Using Fluorescence Cross-Correlation Spectroscopy

    Fluorescence cross-correlation spectroscopy (FCCS) is a promising technique for observing and quantifying protein–protein interactions in vitro and in vivo. FCCS has emerged as a useful tool for obtaining para...

    Manisha Tiwari, Masataka Kinjo in The Nucleus (2015)

  9. No Access

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    Aryl-Acetic and Cinnamic Acids as Lipoxygenase Inhibitors with Antioxidant, Anti-inflammatory, and Anticancer Activity

    Cinnamic acids have been identified as interesting compounds with cytotoxic, anti-inflammatory, and antioxidant properties. Lipoxygenase pathway, catalyzing the first two steps of the transformation of arachid...

    Dimitra Hadjipavlou-Litina, Eleni Pontiki in Advanced Protocols in Oxidative Stress III (2015)

  10. No Access

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    Flow Cytometry Enumeration of Apoptotic Cancer Cells by Apoptotic Rate

    Most authors currently quantify the frequency of apoptotic cells in a given phenotypically defined population after calculating the apoptotic index (AI), i.e., the percentage of apoptotic cells displaying a sp...

    David Diaz B.Sc., Alfredo Prieto B.Sc., Ph.D. in Apoptosis and Cancer (2015)

  11. No Access

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    RNA Fluorescence In Situ Hybridization in Cultured Mammalian Cells

    It is now clear that long noncoding RNAs (lncRNAs) regulate a number of aspects of nuclear organization and gene expression. An important tool for the study of the distribution and function of lncRNAs is RNA f...

    Vidisha Tripathi, Jingyi Fei, Taekjip Ha in Regulatory Non-Coding RNAs (2015)

  12. No Access

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    Quantitative Proteomic Identification of Host Factors Involved in the Salmonella typhimurium Infection Cycle

    Quantitative proteomics, based on stable isotope labeling by amino acids in cell culture (SILAC), can be used to identify host proteins involved in the intracellular interplay with pathogens. This method allow...

    Dora Kaloyanova, Mijke Vogels, Bas W. M. van Balkom, J. Bernd Helms in Salmonella (2015)

  13. No Access

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    Alkylation of Galectin-1 with Iodoacetamide and Mass Spectrometric Mapping of the Sites of Incorporation

    Galectins can display unique sensitivity to oxidative changes that result in significant conformational alterations that prevent carbohydrate recognition. While a variety of approaches can be utilized to preve...

    Sean R. Stowell, Connie M. Arthur, Richard D. Cummings, Christa L. Feasley in Galectins (2015)

  14. No Access

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    Binding and Cytotoxic Trafficking of Cholesterol Hydroperoxides by Sterol Carrier Protein-2

    Redox-active cholesterol hydroperoxides (ChOOHs) generated by oxidative stress in eukaryotic cells may propagate cytotoxic membrane damage by undergoing one-electron reduction or, at low levels, act as mobile ...

    Albert W. Girotti Ph.D., Tamas Kriska in Advanced Protocols in Oxidative Stress III (2015)

  15. No Access

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    Analysis of Receptor Tyrosine Kinase (RTK) Phosphorylation by Immunoblotting

    Immunoblotting for phosphorylated forms of receptor tyrosine kinases (RTKs) has been the mainstay of investigations on RTK signaling for the past two decades. Despite the development of quantitative mass spect...

    Martina McDermott, Norma O’Donovan in Receptor Tyrosine Kinases (2015)

  16. No Access

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    Analysis of the Conductivity of Plasmodesmata by Microinjection

    Pressure microinjection can be used to introduce fluorescent dyes and labeled macromolecules into single cells. The method allows measuring transport activity of macromolecules such as proteins and RNA molecul...

    Friedrich Kragler in Plasmodesmata (2015)

  17. No Access

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    The Use of Experimental Structures to Model Protein Dynamics

    The number of solved protein structures submitted in the Protein Data Bank (PDB) has increased dramatically in recent years. For some specific proteins, this number is very high—for example, there are over 550...

    Ataur R. Katebi, Kannan Sankar, Kejue Jia in Molecular Modeling of Proteins (2015)

  18. No Access

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    Mechanical Nociception Measurement in Mice and Rats with Automated Von Frey Equipment

    Von Frey hairs are important tools for the study of mechanisms of cutaneous stimulation-induced sensory input. Mechanical force is exerted via application of a particular hair to the cutaneous receptive field ...

    Gabriele Campana, Roberto Rimondini in Opioid Receptors (2015)

  19. No Access

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    Wheat (Triticum aestivum L.) Transformation Using Immature Embryos

    Wheat may now be transformed very efficiently by Agrobacterium tumefaciens. Under the protocol hereby described, immature embryos of healthy plants of wheat cultivar Fielder grown in a well-conditioned greenhouse...

    Yuji Ishida, Masako Tsunashima, Yukoh Hiei, Toshihiko Komari in Agrobacterium Protocols (2015)

  20. No Access

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    Measurement of Macrophage Toll-Like Receptor 4 Expression After Morphine Treatment

    The immune system is a complex and finely orchestrated system, and many soluble molecules and receptors contribute to its regulation.

    Mara Castelli, Alberto Panerai, Paola Sacerdote, Silvia Franchi in Opioid Receptors (2015)

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