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    In vivo Live Imaging of Calcium Waves and Other Cellular Processes during Fertilization in Caenorhabditis elegans
    Authors:  Jun Takayama, Masashi Fujita and Shuichi Onami, date: 04/05/2017, view: 1895, Q&A: 0
    [Abstract] Fertilization calcium waves are a conserved trigger for animal development; however, genetic analysis of these waves has been limited due to the difficulty of imaging in vivo fertilization. Here we describe a protocol to image calcium dynamics during in vivo fertilization in the genetic animal model Caenorhabditis elegans ...
    Two-electrode Voltage-clamp Recordings in Xenopus laevis Oocytes:Reconstitution of Abscisic Acid Activation of SLAC1 Anion Channel via PYL9 ABA Receptor
    Authors:  Cun Wang, Jingbo Zhang and Julian I. Schroeder, date: 01/20/2017, view: 2041, Q&A: 0
    [Abstract] Two-Electrode Voltage-Clamp (TEVC) recording in Xenopus laevis oocytes provides a powerful method to investigate the functions and regulation of ion channel proteins. This approach provides a well-known tool to characterize ion channels or transporters expressed in Xenopus laevis oocytes. The plasma membrane of the oocyte is ...
    Measurement of H+ Flux in Rice by Non-invasive Micro-test Technology
    Authors:  Xi Zhang, Xiu-Yue Zhang and Li-Ping Yin, date: 12/05/2015, view: 2265, Q&A: 0
    [Abstract] Rice plants release proton (H+) from root cells into rhizosphere area leading to the acidification of the rhizosphere and increased solubility of ferric iron complexes on the cell membrane, which is important for iron uptakes. Here, we present a detailed protocol to measure H+ flux in root hairs of transgenic rice seedlings ...
    An Assay to Test the Capacity of Arabidopsis Plant Defensin Type1 Protein to Induce Cellular Zinc (Zn) Tolerance in Yeast
    Authors:  Zaigham Shahzad, Laurence Marquès, Pierre Berthomieu and Françoise Gosti, date: 11/20/2015, view: 2566, Q&A: 0
    [Abstract] Heterologous expression of genes in budding yeast Saccharomyces cerevisiae (S. cerevisiae) is especially suitable to functionally study the corresponding encoded protein at the cellular level (Bonneaud et al., 1991). This is mainly because many strains defective in specific activities are available and could be ...
    Loading of Cells with Fluorescent Probe to Study Intracellular Acid-base Homeostasis in Lactic Acid Bacteria
    Authors:  Pablo Mortera, Federico Zuljan, Christian Magni and Sergio H. Alarcón, date: 01/20/2015, view: 4308, Q&A: 0
    [Abstract] Here we describe a protocol which we have used to study the homeostasis intracellular in vivo in lactic acid bacteria (LAB) using a fluorescent probe. This type of probes can be used for determining changes in the pH of cytoplasm with high sensitivity, temporal resolution and technical simplicity as well as accessing the rate of change ...
    A Protocol for Measurement of Intracellular pH
    Authors:  Iman Saramipoor Behbahan, Matthew A. McBrian and Siavash K. Kurdistani, date: 01/20/2014, view: 8046, Q&A: 1
    [Abstract] Intracellular pH (pHi) is an important physiological determinant of enzyme activity and cellular function (Kurkdjian and Guern, 1989). All proteins depend on a tightly regulated pH to maintain their structure and function. Protonation–deprotonation events can dictate the charge of biological surfaces and are integral steps in many ...
    Transport Assays in Aspergillus nidulans
    Authors:  Emilia Krypotou and George Diallinas, date: 11/20/2013, view: 3370, Q&A: 0
    [Abstract] Transport assays allow the direct kinetic analysis of a specific transporter by measuring apparent Km and Vmax values, and permit the characterization of substrate specificity profiles through competition assays. In this protocol, we describe a rapid and easy method for performing uptake assays in the model ...
    Measurement of Extracellular Ca2+ Influx and Intracellular H+ Efflux in Response to Glycerol and PEG6000 Treatments
    Authors:  Tao Li and Baodong Chen, date: 09/20/2013, view: 4175, Q&A: 0
    [Abstract] The characteristics of Ca2+ and H+ fluxes may reflect the activities of aquaporins, as the up-regulation of aquaporin activities is directly associated with the decrease in cytoplasmic H+ concentration and increase in cytoplasmic Ca2+ concentration. The higher aquaporin activities can protect cells ...
    Flow Cytometric Analysis of Calcium Influx Assay in T cells
    Author:  Sun-Hwa Lee, date: 09/20/2013, view: 7483, Q&A: 0
    [Abstract] Calcium influx is one of the key signaling events upon stimulation of T cell receptors (TCR) and plays an important role for T cell activation, proliferation, and differentiation. Phorbol myristate acetate (PMA) and calcium ionophore ionomycin are commonly utilized as stimulants in a variety of immunologic assays including T cell activation. PMA ...
    Sodium Current Measurements in HEK293 Cells
    [Abstract] This protocol is used to measure sodium currents from heterologously transfected cells lines, such as HEK293 cells. Standard whole cell patch clamp technique is used to assess ion channel function. This protocol has been used to study cardiac type Nav1.5 sodium channel, and in particular to compare wild-type and mutant channels related ...