Mammalia

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    Protocols in Current Issue
    Quantifying HIV-1-Mediated Gut CD4+ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model
    [Abstract] Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture (LPAC) model. The LPAC model is ideal for studying CD4 T cell death induced by clinically-relevant ...
    Ribonucleoprotein Immunoprecipitation (RIP) Analysis
    Authors:  Jennifer L. Martindale, Myriam Gorospe and Maria L. Idda, date: 01/20/2020, view: 468, Q&A: 0
    [Abstract] RNAs and RNA-binding proteins (RBPs) can interact dynamically in ribonucleoprotein (RNP) complexes that play important roles in controlling gene expression programs. One of the powerful ways to investigate changes in the association of RNAs with an RBP of interest is by immunoprecipitation (IP) analysis of native RNPs. RIP (RNP ...
    cDNA Display Mediated Immuno-PCR (cD-IPCR): A Novel PCR-based Antigen Detection Method
    Authors:  Chathuni Jayathilake, Takuya Terai and Naoto Nemoto, date: 12/20/2019, view: 715, Q&A: 0
    [Abstract] Immuno-PCR (IPCR) is a powerful method in antigen detection where a PCR-amplifiable DNA reporter is conjugated to a specific antibody or an aptamer for the target molecule. In the development and application of IPCR, successful conjugation of a protein (an antibody) with a reporter DNA becomes challenging. To address this issue, we recently ...
    Characterization of Biological Motion Using Motion Sensing Superpixels
    [Abstract] Precise spatiotemporal regulation is the foundation for the healthy development and maintenance of living organisms. All cells must correctly execute their function in the right place at the right time. Cellular motion is thus an important dynamic readout of signaling in key disease-relevant molecular pathways. However despite the rapid ...
    A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material
    [Abstract] We describe here a detailed, refined protocol for the generation of citrulline-specific monoclonal antibodies from single human B cells from rheumatoid arthritis (RA) patients. This protocol provides a detailed guide of the procedure starting from single B cells of your choice and followed by amplification of the variable region of immunoglobulin ...
    In vitro Infection of Primary Human Monocytes with HIV-1
    Authors:  Patrick O’Connell, Yong-Hui Zheng, Andrea Amalfitano and Yasser A. Aldhamen, date: 07/05/2019, view: 1145, Q&A: 0
    [Abstract] Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 ...
    TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4+ T Cells
    Authors:  Anwesha Sanyal, Vatsala S. Rangachar and Phalguni Gupta, date: 05/20/2019, view: 2131, Q&A: 1
    [Abstract] The latent HIV-1 viral reservoir in resting CD4+ (rCD4+) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the ...
    Assessment of Humoral Alloimmunity in Mixed Lymphocyte Reaction
    Authors:  Georgios Pissas and Theodoros Eleftheriadis, date: 01/20/2019, view: 2916, Q&A: 1
    [Abstract] Humoral alloimmunity remains a significant and unresolved problem that constrains allograft survival. Thus, there is a need for the development of an easy, preferably non-radioactive, and inexpensive protocol for assessing the effect of various drug treatments on humoral alloimmunity. In order to satisfy this demand, we developed such a protocol ...
    Monitoring Natural Killer Cell Function in Human Ovarian Cancer Cells of Ascitic Fluid
    Authors:  Vikash Kumar and Asok Mukhopadhyay, date: 12/20/2018, view: 2011, Q&A: 0
    [Abstract] Natural killer (NK) cells are the major effectors of the innate immune system when activated resulting in modulation of immune response of the host defense through target cell lysis and secretion of cytokines. Precise functions of NK cells are essential for the treatment outcome of different virus infections and malignant diseases. NK cells impart ...
    Quantitation of Regulatory Activity for the Complement Alternative Pathway Using an Adaptation of the AP50 in vitro Assay
    Authors:  Sheila Cabezas Falcon, David L Gordon and Jillian M Carr, date: 12/20/2018, view: 1719, Q&A: 2
    [Abstract] Complement pathways function to identify and remove pathogens and infected cells. There are three complement pathways: the classical, lectin and alternative pathway (AP). While all pathways are activated following pathogen stimuli, the AP is constitutively active and tightly controlled by activators (e.g., Factor B, Factor D) and negative ...



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