Immunology

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    Protocols in Current Issue
    Plasmodium cynomolgi Berok Growth Inhibition Assay by Thiol-reactive Probe Based Flow Cytometric Measurement
    Authors:  Jessica Jie Ying Ong, Bruce Russell and Jin-Hee Han, date: 09/05/2021, view: 609, Q&A: 0

    The relapsing malaria species, Plasmodium vivax, is the most widely distributed and difficult-to-treat cause of human malaria. The merozoites of P. vivax preferentially invade ephemeral human CD71+ reticulocytes (nascent reticulocytes), thereby

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    Isolation and Quantification of Mouse γδT-cells in vitro and in vivo
    Authors:  Isha Rana, Krithika Badarinath, Ravindra K. Zirmire and Colin Jamora, date: 09/05/2021, view: 725, Q&A: 0

    The skin plays an important role in protecting the body from pathogens and chemicals in the external environment. Upon injury, a healing program is rapidly initiated and involves extensive intercellular communication to restore tissue homeostasis.

    ...
    Microscopic Detection of ASC Inflammasomes in Bone Marrow Derived Macrophages Post Stimulation
    Authors:  Longjun Li, Rudi Mao and Xiaopeng Qi, date: 09/05/2021, view: 629, Q&A: 0

    An inflammasome is an intracellular multiprotein complex that plays important roles in host defense and inflammatory responses. Inflammasomes are typically composed of the adaptor protein apoptosis-associated speck-like protein containing a CARD

    ...
    Plasmodium cynomolgi Berok Growth Inhibition Assay by Thiol-reactive Probe Based Flow Cytometric Measurement
    Authors:  Jessica Jie Ying Ong, Bruce Russell and Jin-Hee Han, date: 09/05/2021, view: 609, Q&A: 0
    [Abstract]

    The relapsing malaria species, Plasmodium vivax, is the most widely distributed and difficult-to-treat cause of human malaria. The merozoites of P. vivax preferentially invade ephemeral human CD71+ reticulocytes (nascent reticulocytes), thereby limiting the development of a robust continuous culture in vitro. Fortunately, P. vivax’s sister

    ...
    Isolation and Quantification of Mouse γδT-cells in vitro and in vivo
    Authors:  Isha Rana, Krithika Badarinath, Ravindra K. Zirmire and Colin Jamora, date: 09/05/2021, view: 725, Q&A: 0
    [Abstract]

    The skin plays an important role in protecting the body from pathogens and chemicals in the external environment. Upon injury, a healing program is rapidly initiated and involves extensive intercellular communication to restore tissue homeostasis. The deregulation of this crosstalk can lead to abnormal healing processes and is the foundation of

    ...
    Microscopic Detection of ASC Inflammasomes in Bone Marrow Derived Macrophages Post Stimulation
    Authors:  Longjun Li, Rudi Mao and Xiaopeng Qi, date: 09/05/2021, view: 629, Q&A: 0
    [Abstract]

    An inflammasome is an intracellular multiprotein complex that plays important roles in host defense and inflammatory responses. Inflammasomes are typically composed of the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), cytoplasmic sensor protein, and the effector protein pro-caspase-1. ASC assembly into a protein

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    Construction of a Highly Diverse mRNA Library for in vitro Selection of Monobodies
    [Abstract]

    Recently, we developed transcription/translation coupled with the association of puromycin linker (TRAP) display as a quick in vitro selection method to obtain antibody-like proteins. For the in vitro selection, it is important to prepare mRNA libraries among which the diversity is high. Here, we describe a method for the preparation of monobody

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    Preparation of Drosophila Larval Blood Cells for Single-cell RNA Sequencing
    Authors:  Sudhir Gopal Tattikota and Norbert Perrimon, date: 08/20/2021, view: 466, Q&A: 0
    [Abstract]

    Recent advances in single-cell RNA-sequencing (scRNA-seq) technologies provide unprecedented opportunities to identify new cell types and characterize cell states. One of the most important requirements for performing scRNA-seq is to obtain high-quality single cells in suspension. Recently, we used this approach to characterize Drosophila blood

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    One-step White Blood Cell Extracellular Staining Method for Flow Cytometry
    [Abstract]

    Flow cytometry is a powerful analytical technique that is increasingly used in scientific investigations and healthcare; however, it requires time-consuming, multi-step sample procedures, which limits its use to specialized laboratories. In this study, we propose a new universal one-step method in which white blood cell staining and red blood cell

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    Isolation of Microglia and Analysis of Protein Expression by Flow Cytometry: Avoiding the Pitfall of Microglia Background Autofluorescence
    Authors:  Jeremy C. Burns, Richard M. Ransohoff and Michaël Mingueneau, date: 07/20/2021, view: 1444, Q&A: 0
    [Abstract]

    Microglia are a unique type of tissue-resident innate immune cell found within the brain, spinal cord, and retina. In the healthy nervous system, their main functions are to defend the tissue against infectious microbes, support neuronal networks through synapse remodeling, and clear extracellular debris and dying cells through phagocytosis. Many

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    Production, quantification, and infection of Amazonian Phlebovirus (Bunyaviridae)
    [Abstract]

    Phlebotomine vectors, sand flies of the order Diptera, are known to transmit Leishmania parasites as well as RNA viruses (arboviruses) to humans. The arbovirus, Icoaraci Phlebovirus (BeAN 24262 - ICOV), used in this study was isolated from Nectomys rodents, a mammalian species that is the same natural sylvatic reservoir of Leishmania (Leishmania)

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    A Lipidomics Approach to Measure Phosphatidic Acid Species in Subcellular Membrane Fractions Obtained from Cultured Cells
    Authors:  Nawal Kassas, Laetitia Fouillen, Stéphane Gasman and Nicolas Vitale, date: 06/20/2021, view: 1131, Q&A: 0
    [Abstract]

    Over the last decade, lipids have emerged as possessing an ever-increasing number of key functions, especially in membrane trafficking. For instance, phosphatidic acid (PA) has been proposed to play a critical role in different steps along the secretory pathway or during phagocytosis. To further investigate in detail the precise nature of PA

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    Multi-color Flow Cytometry for Comprehensive Analysis of the Tumor Immune Infiltrate in a Murine Model of Breast Cancer
    Authors:  Ana S. Almeida, Miriam R. Fein and Mikala Egeblad, date: 06/05/2021, view: 1668, Q&A: 0
    [Abstract]

    Flow cytometry is a popular laser-based technology that allows the phenotypic and functional characterization of individual cells in a high-throughput manner. Here, we describe a detailed procedure for preparing a single-cell suspension from mammary tumors of the mouse mammary tumor virus-polyoma middle T (MMTV-PyMT) and analyzing these cells by

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