Immunology

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    Protocols in Current Issue
    Quantification of Serum Ovalbumin-specific Immunoglobulin E Titre via in vivo Passive Cutaneous Anaphylaxis Assay
    [Abstract] Murine models of allergic airway disease are frequently used as a tool to elucidate the cellular and molecular mechanisms of tissue-specific asthmatic disease pathogenesis. Paramount to the success of these models is the induction of experimental antigen sensitization, as indicated by the presence of antigen-specific serum immunoglobulin E. The ...
    Characterization of Mouse Adult Testicular Macrophage Populations by Immunofluorescence Imaging and Flow Cytometry
    Authors:  Noushin Mossadegh-Keller and Michael H. Sieweke, date: 03/05/2019, view: 1143, Q&A: 0
    [Abstract] Testicular macrophages (tMΦ) are the most abundant immune cells residing in the testis, an immune-privileged organ. TMΦ are known to exhibit different functions, such as protecting spermatozoa from auto-immune attack by producing immunosuppressive cytokines and trophic roles in supporting spermatogenesis and male sex hormone production. They also ...
    DNA Immunization Using in vivo Electroporation for Generating Monoclonal Antibodies Against Mouse IL-9R
    [Abstract] Membrane proteins such as cytokine receptors and G protein-coupled receptors can be drug targets. Recently, we have generated specific monoclonal antibodies (mAbs) against the mouse IL-9 receptor (IL-9R) and found that IL-9R on memory B cells have critical roles in T-dependent immune response. So far, most antibodies against cell surface proteins ...
    Electroporation of Labeled Antibodies to Visualize Endogenous Proteins and Posttranslational Modifications in Living Metazoan Cell Types
    Authors:  Sascha Conic, Dominique Desplancq, László Tora and Etienne Weiss, date: 11/05/2018, view: 1564, Q&A: 0
    [Abstract] The spatiotemporal localization of different intracellular factors in real-time and their detection in live cells are important parameters to understand dynamic protein-based processes. Therefore, there is a demand to perform live-cell imaging and to measure endogenous protein dynamics in single cells. However, fluorescent labeling of endogenous ...
    Qualitative and Quantitative Assay for Detection of Circulating Autoantibodies against Human Aortic Antigen
    Authors:  Brent Veerman and Ritu Chakravarti, date: 07/05/2017, view: 4728, Q&A: 0
    [Abstract] Increased amount of autoantibodies in human sera are the hallmark of autoimmune diseases (Wang et al., 2015). In case of known antigen, detection of autoantibodies is done using laboratory based methods. However, in most autoimmune diseases, knowledge of self-antigen is still vague. We have developed an ELISA-based quantitative assay to ...
    Coupling of HIV-1 gp120-derived Core Protein to Paramagnetic Beads and Adsorption Assays
    Authors:  Jidnyasa Ingale and Richard T Wyatt, date: 10/05/2015, view: 4438, Q&A: 0
    [Abstract] Analysis of the functional activity in polyclonal serum following immunization of a complex protein or glycoprotein immunogen is a very important but tedious process. Fine mapping of epitope-specific antibodies is difficult when they are elicited at relatively low levels. In our recent study focused on developing an HIV-1 vaccine, we immunized ...
    Pulse Chase of Suspension Cells
    Authors:  Lai-Yee Wong, QiMing Liang, Kevin Brulois and Jae Jung, date: 07/05/2014, view: 8338, Q&A: 0
    [Abstract] Pulse-chase method is a powerful technique used to follow the dynamics of proteins over a period of time. The expression level, processing, transport, secretion or half-life of proteins can be tracked by metabolically labeling the cells, such as with radiolabeled amino acids (pulse step). This protocol describes the condition used to study the ...
    Monoclonal Antibody Purification (Nicotiana benthamiana Plants)
    Authors:  Adam Husk, Krystal Teasley Hamorsky and Nobuyuki Matoba, date: 01/20/2014, view: 9527, Q&A: 1
    [Abstract] Plant-based expression systems provide an alternative biomanufacturing platform for recombinant proteins (Matoba et al., 2011). In particular, plant virus-based vectors can overexpress proteins within days in the leaf tissue of Nicotiana benthamiana (N. benthamiana). To overcome the issues of genetic instability and ...
    Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-cardiolipin Antibodies
    Author:  Zheng Liu, date: 06/20/2012, view: 9416, Q&A: 0
    [Abstract] The circulating anticardiolipin antibody is a hallmark of antiphospholipid syndrome. It also appears in a number of autoimmune mouse models and is indicative of the break of tolerance against self antigens. This protocol describes a reliable method to determine the relative serum titer of anticardiolipin in autoimmune mouse models.
    Isolation of Circulating Immune Complexes from TB Patient Serum for Serodiagnosis
    Authors:  Uma Devi Ranganathan, Ramalingam Bethunaickan and Alamelu Raja, date: 06/05/2012, view: 10226, Q&A: 0
    [Abstract] Estimation of circulating immune complex in tuberculosis patients has shown better insight to the infection. Isolating circulating immune complex helps quantifying both antigen and antibody in the serum. It’s a simple procedure to improve the sensitivity and specificity in serodiagnosis of tuberculosis. This protocol may be modified to detect ...



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