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    Protocols in Current Issue
    Simple Method to Determine Protein Redox State in Arabidopsis thaliana
    Authors:  Keisuke Yoshida and Toru Hisabori, date: 06/05/2019, view: 1243, Q&A: 0
    [Abstract] Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...
    Quantification of the Humidity Effect on HR by Ion Leakage Assay
    Authors:  Musoki Mwimba and Xinnian Dong, date: 04/05/2019, view: 996, Q&A: 0
    [Abstract] We describe a protocol to measure the contribution of humidity on cell death during the effector-triggered immunity (ETI), the plant immune response triggered by the recognition of pathogen effectors by plant resistance genes. This protocol quantifies tissue cell death by measuring ion leakage due to loss of membrane integrity during the ...
    Visualization of Plant Cell Wall Epitopes Using Immunogold Labeling for Electron Microscopy
    Author:  Mateusz Majda, date: 04/05/2019, view: 1326, Q&A: 0
    [Abstract] Plant cell walls consist of different polysaccharides and structural proteins, which form a rigid layer located outside of the plasma membrane. The wall is also a very dynamic cell composite, which is characterized by complex polysaccharide interactions and various modifications during cell development. The visualization of cell wall components in ...
    Quantitative Plasmodesmata Permeability Assay for Pavement Cells of Arabidopsis Leaves
    Authors:  Min Diao, Qiannan Wang and Shanjin Huang, date: 04/05/2019, view: 1364, Q&A: 0
    [Abstract] Plasmodesmata (PD) are intercellular channels between walled plant cells that enable the transportation of materials between adjacent cells, which are important for plant growth and development. The permeability of PD must be tightly regulated. Assays to determine the permeability of PD are crucial for related studies on the regulation of PD ...
    Detection of Disulfides in Protein Extracts of Arabidopsis thaliana Using Monobromobimane (mBB)
    Authors:  Shin‐nosuke Hashida and Maki Kawai-Yamada, date: 03/05/2019, view: 1230, Q&A: 0
    [Abstract] Thiol-disulfide exchange is a key posttranslational modification, determining the folding process of intra- and inter-protein structures. Thiols can be detected by colorimetric reagents, which are stoichiometrically reduced by free thiols, and by fluorescent adducts, showing fluorescence only after thioester formation. We adapted a simple ...
    Visualization and Quantification of Cell-to-cell Movement of Proteins in Nicotiana benthamiana
    Authors:  Mila C Blekemolen, Mariliis Tark-Dame and Frank L.W Takken, date: 12/20/2018, view: 1901, Q&A: 0
    [Abstract] Cell-to-cell movement of proteins through plasmodesmata is a widely-established mechanism for intercellular signaling in plants. Current techniques to study intercellular protein translocation rely on single-cell transformation using particle bombardment or transgenic lines expressing photo-inducible fluorophores. The method presented here allows ...
    Using Arabidopsis Mesophyll Protoplasts to Study Unfolded Protein Response Signaling
    Authors:  Yan Bao and Diane C. Bassham, date: 12/05/2018, view: 1830, Q&A: 0
    [Abstract] Various environmental stresses or artificial reagents can trigger unfolded protein accumulation in the endoplasmic reticulum (ER) due to the folding capacity of the ER being exceeded. This is termed ER stress, and triggers the unfolded protein response (UPR). Assays for activation of the UPR in plants include Tunicamycin (Tm)- or dithiothreitol ...
    In planta Transcriptome Analysis of Pseudomonas syringae
    Authors:  Tatsuya Nobori and Kenichi Tsuda, date: 09/05/2018, view: 3042, Q&A: 0
    [Abstract] Profiling bacterial transcriptome in planta is challenging due to the low abundance of bacterial RNA in infected plant tissues. Here, we describe a protocol to profile transcriptome of a foliar bacterial pathogen, Pseudomonas syringae pv. tomato DC3000, in the leaves of Arabidopsis thaliana at an early stage of ...
    Isothermal Titration Calorimetry: A Biophysical Method to Characterize the Interaction between Label-free Biomolecules in Solution
    Author:  Andrea Saponaro, date: 08/05/2018, view: 8799, Q&A: 0
    [Abstract] This protocol can be applied to analyze the direct interaction between a soluble protein and a target ligand molecule using Isothermal Titration Calorimetry (ITC, Malvern). ITC allows the biophysical characterization of binding between label-free, non-immobilized and in-solution biomolecules by providing the stoichiometry of the interaction, the ...
    Quantification of Starch in Guard Cells of Arabidopsis thaliana
    Authors:  Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 3307, 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.



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