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    Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
    Authors:  Steffen Mayerl and Charles ffrench-Constant, date: 01/05/2021, view: 392, Q&A: 0
    [Abstract]

    The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain

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    A Simple Microplate Assay for Reactive Oxygen Species Generation and Rapid Cellular Protein Normalization
    Authors:  Neville S. Ng and Lezanne Ooi, date: 01/05/2021, view: 711, Q&A: 0
    [Abstract] 2',7'-dichlorofluorescein (DCF) and derivatives are commonly used as fluorescent indicators of a broad spectrum of reactive oxygen species (ROS) generation in cell-based assays. However, there are numerous challenges inherent to the utilization of DCF probes for intracellular microscopic analysis, including bleaching and probe efflux. Plate ...
    A Transient Transfection-based Cell Adhesion Assay with 293T Cells
    Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 513, Q&A: 0
    [Abstract]

    The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a transient 293T cell transfection-based cell adhesion assay to study selective cell adhesion of a

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    A Quantitative Assay to Measure Stress Granule Association of Proteins and Peptides in Semi-permeabilized Human Cells
    Authors:  Saskia Hutten and Dorothee Dormann, date: 12/20/2020, view: 828, Q&A: 0
    [Abstract]

    Stress granules (SGs) are membrane-less organelles that form in the cytoplasm through phase separation, in response to diverse stressors. SGs contain translationally stalled mRNAs, proteins involved in translation, and various RNA-binding proteins (RBPs). Due to the high local concentration of aggregation-prone RBPs, SGs might act as condensation

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    RNA ImmunoGenic Assay: A Method to Detect Immunogenicity of in vitro Transcribed mRNA in Human Whole Blood
    [Abstract] The mRNA therapeutics is a new class of medicine to treat many various diseases. However, in vitro transcribed (IVT) mRNA triggers immune responses due to recognition by human endosomal and cytoplasmic RNA sensors, but incorporation of modified nucleosides have been shown to reduce such responses. Therefore, an assay signifying important ...
    Equilibrium and Kinetic Measurements of Ligand Binding to HiBiT-tagged GPCRs on the Surface of Living Cells
    Authors:  Michelle E. Boursier, Sergiy Levin, Robin Hurst and Rachel Friedman Ohana, date: 12/20/2020, view: 565, Q&A: 0
    [Abstract]

    G-protein coupled receptors (GPCRs) remain at the forefront of drug discovery efforts. Detailed assessment of features contributing to GPCR ligand engagement in a physiologically relevant environment is imperative to the development of new therapeutics with improved efficacy. Traditionally, binding properties such as affinity and kinetics were

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    Native Co-immunoprecipitation Assay to Identify Interacting Partners of Chromatin-associated Proteins in Mammalian Cells
    Authors:  Afzal Husain, Nasim A. Begum, Maki Kobayashi and Tasuku Honjo, date: 12/05/2020, view: 972, Q&A: 0
    [Abstract]

    Protein-protein interactions play key roles in nuclear processes including transcription, replication, DNA damage repair, and recombination. Co-immunoprecipitation (Co-IP) followed by western blot or mass spectrometry is an invaluable approach to identify protein-protein interactions. One of the challenges in the Co-IP of a protein localized to

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    Generating Three-dimensional Human Granulomas in vitro to Study Mycobacterium tuberculosis-host Interaction
    Authors:  Ainhoa Arbués, Michael Kammüller and Damien Portevin, date: 11/20/2020, view: 690, Q&A: 0
    [Abstract] Granulomas are organized multicellular structures that constitute the hallmark of an infection by the human pathogen Mycobacterium tuberculosis (Mtb). A better understanding of the complex host-Mtb interactions within the granuloma’s environment may lead to new therapeutic or preventive tools to improve the control of the ...
    Confocal Microscopy of Reovirus Transport in Living Dorsal Root Ganglion Neurons
    Authors:  Pavithra Aravamudhan, Krishnan Raghunathan and Terence S. Dermody, date: 11/20/2020, view: 654, Q&A: 0
    [Abstract]

    Neurotropic reoviruses repurpose host machinery to traffic over long distances in neuronal processes and access distal replication sites. Understanding mechanisms of neuronal transmission is facilitated by using simplified in vitro primary neuronal culture models. Advances in the design of compartmentalized microfluidic devices lend robustness to

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    K+ Release Assay and K+ Measurement in Oocyte Assay
    Author:  Hong Li, date: 11/05/2020, view: 718, Q&A: 0
    [Abstract] The Xenopus oocyte is a powerful system for the exogenous expression and functional characterization of plant membrane transport proteins. Until now, a number of potassium transporters and channels have been identified in oocytes expression system by the two-electrode voltage clamp technology. It is difficult to characterize K+ ...



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