Cell Biology

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    Protocols in Current Issue
    Electroporation of Labeled Antibodies to Visualize Endogenous Proteins and Posttranslational Modifications in Living Metazoan Cell Types
    Authors:  Sascha Conic, Dominique Desplancq, László Tora and Etienne Weiss, date: 11/05/2018, view: 1884, Q&A: 0
    [Abstract] The spatiotemporal localization of different intracellular factors in real-time and their detection in live cells are important parameters to understand dynamic protein-based processes. Therefore, there is a demand to perform live-cell imaging and to measure endogenous protein dynamics in single cells. However, fluorescent labeling of endogenous ...
    Transcytosis Assay for Transport of Glycosphingolipids across MDCK-II Cells
    Authors:  Maria Daniela Garcia-Castillo, Wayne I. Lencer and Daniel J.-F. Chinnapen, date: 10/20/2018, view: 1953, Q&A: 0
    [Abstract] Absorption and secretion of peptide and protein cargoes across single-cell thick mucosal and endothelial barriers occurs by active endocytic and vesicular trafficking that connects one side of the epithelial or endothelial cell (the lumen) with the other (the serosa or blood). Assays that assess this pathway must robustly control for non-specific ...
    Boron Uptake Assay in Xenopus laevis Oocytes
    Authors:  Sheliang Wang, Namiki Mitani-Ueno and Junpei Takano, date: 03/05/2018, view: 3128, Q&A: 0
    [Abstract] Boron (B) is essential for plant growth and taken up by plant roots as boric acid. Under B limitation, B uptake and translocation in plants are dependent on the boric acid channels located in the plasma membrane. Xenopus leavis oocyte is a reliable heterologous expression system to characterize transport activities of boric acid channels ...
    Microvesicle Isolation from Rat Brain Extract Treated Human Mesenchymal Stem Cells
    Authors:  Ji Yong Lee, Seong-Mi Choi and Han-Soo Kim, date: 07/05/2017, view: 5769, Q&A: 0
    [Abstract] Microvesicle (MVs) are submicron-sized membranous vesicles that are either actively released from cells via secretory compartments or shed from cell surface membranes. MVs are generated by many cell types and serve as vehicles that transfer biological information (e.g., protein, mRNA, and miRNA) to distant cells, thereby affecting their ...
    Analysis of Mitochondrial Transfer in Direct Co-cultures of Human Monocyte-derived Macrophages (MDM) and Mesenchymal Stem Cells (MSC)
    Authors:  Megan V. Jackson and Anna D. Krasnodembskaya, date: 05/05/2017, view: 6829, Q&A: 0
    [Abstract] Mesenchymal stem/stromal cells (MSC) are adult stem cells which have been shown to improve survival, enhance bacterial clearance and alleviate inflammation in pre-clinical models of acute respiratory distress syndrome (ARDS) and sepsis. These diseases are characterised by uncontrolled inflammation often underpinned by bacterial infection. The ...
    In vitro Assays for Measuring Endothelial Permeability by Transwells and Electrical Impedance Systems
    Authors:  Hong-Ru Chen and Trai-Ming Yeh, date: 05/05/2017, view: 13744, Q&A: 1
    [Abstract] Vascular leakage is an important feature in several diseases, such as septic shock, viral hemorrhagic fever, cancer metastasis and ischemia-reperfusion injuries. Thus establishing assays for measuring endothelial permeability will provide insight into the establishment or progression of such diseases. Here, we provide transwell permeability assay ...
    Secretion of Adipsin as an Assay to Measure Flux from the Endoplasmic Reticulum (ER)
    Authors:  Alexandria Brumfield, Natasha Chaudhary and Timothy E. McGraw, date: 04/05/2017, view: 4335, Q&A: 0
    [Abstract] In this protocol we describe a quantitative biochemical assay to assess the efficiency of endoplasmic reticulum (ER) to Golgi protein transport in adipocytes (Bruno et al., 2016). The assay takes advantage of the fact that adipocytes secrete various bioactive proteins, known as adipokines. As a measure of ER to Golgi flux we determine the ...
    Uptake Assay for Radiolabeled Peptides in Yeast
    Authors:  Melinda Hauser, Houjian Cai, Fred Naider and Jeffrey M. Becker, date: 11/20/2016, view: 4937, Q&A: 0
    [Abstract] We describe an assay for measuring the uptake of radioactive peptides into the yeast Saccharomyces cerevisiae. The methods presented here can be adapted to measure a variety of substrates transported into any bacterial or fungal cell via specific carrier-mediated systems.
    Halo Assay for Toxic Peptides and Other Compounds in Microorganisms
    Authors:  Houjian Cai, Melinda Hauser, Fred Naider and Jeffrey M. Becker, date: 11/20/2016, view: 5177, Q&A: 0
    [Abstract] We describe an assay for determination of toxicity in S. cerevisiae involving spotting of a toxic peptide on a lawn of yeast cells. This assay may be generalized to determine toxicity of a variety of compounds by substituting a putative toxic compound in place of the peptide. The general protocol may also be used to determine toxicity of ...
    VAMP8-3xHA Uptake Assay in HeLa Cells
    Authors:  Steve Jean and Amy A. Kiger, date: 02/20/2016, view: 4729, Q&A: 0
    [Abstract] Transmembrane proteins are rarely exclusively localized to a specific vesicle or an organelle. Most transmembrane proteins undergo complicated trafficking routes. Thus, transmembrane proteins are under constant flux, and at steady state, found on a variety of vesicles or organelles. This characteristic makes the study of their trafficking routes ...



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