Protocols in Current Issue
    An Image-based Dynamic High-throughput Analysis of Adherent Cell Migration

    In this protocol, we describe a method to monitor cell migration by live-cell imaging of adherent cells. Scratching assay is a common method to investigate cell migration or wound healing capacity. However, achieving homogenous scratching, finding the optimal time window for end-point analysis and performing an objective image analysis imply, even

    Multiplication and Growth Inhibition Activity Assays for the Zoonotic Malaria Parasite, Plasmodium knowlesi
    Authors:  Franziska Mohring, Thomas A. Rawlinson, Simon J. Draper and Robert W. Moon, date: 09/05/2020, view: 2340, Q&A: 0
    [Abstract] Malaria remains a major cause of morbidity and mortality globally. Clinical symptoms of the disease arise from the growth and multiplication of Plasmodium parasites within the blood of the host. Thus in vitro assays to determine how drug, antibody and genetic perturbations affect the growth rate of Plasmodium parasites ...
    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 ...
    Nab Escaping AAV Mutants Isolated from Mouse Muscles
    Authors:  Zheng Chai, R. Jude Samulski and Chengwen Li, date: 05/05/2018, view: 4632, Q&A: 0
    [Abstract] Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These ...
    Hemagglutination Inhibition (HI) Assay of Influenza Viruses with Monoclonal Antibodies
    [Abstract] Heamagglutination is inhibited when antibodies are present because antibodies to the influenza virus will prevent attachment of the virus to red blood cells. The highest dilution of antibody that prevents hemagglutination is called the HI titer. Human monoclonal antibodies generated from single human B cells were tested to characterize their ...
    Micro Neutralization (MN) Assay of Influenza Viruses with Monoclonal Antibodies
    [Abstract] The human monoclonal antibodies generated from single human B cells were tested to characterize their ability to neutralize virus infectivity. The microneutralization assay is a highly sensitive and specific assay for detecting virus-specific neutralizing antibodies to influenza viruses. This protocol is to measure the ability of human monoclonal ...
    Cell-based Assays to Monitor AID Activity
    [Abstract] The enzyme Activation induced deaminase (AID) underpins antibody affinity maturation and isotype switching through its mutagenic activity of deaminating deoxycytidine to deoxyuridine in DNA. Subsequent processing of the deoxyuridine initiates the processes of somatic hypermutation (SHM) and class switch recombination (CSR) in B cells. ...
    Binding Affinity Measurement of Antibodies from Crude Hybridoma Samples by SPR
    [Abstract] Surface Plasmon Resonance (SPR) is widely used to generate kinetic and affinity information on specific interactions between biomolecules. This technique is label-free and monitors the binding event in real-time. It is generally used for characterization of monoclonal antibody - antigen interactions. This protocol describes specifically the use of ...
    Detection of Tumor Cell Surface-reactive Antibodies
    Authors:  Brian M. Andersen, Michelle R. Goulart, Michael R. Olin and G. Elizabeth Pluhar, date: 05/05/2014, view: 6844, Q&A: 0
    [Abstract] Vaccine-based immunotherapy is being used to treat dogs with primary brain tumors. The vaccines are composed of a lysate of autologous tumor cells, which stimulate an immune response producing tumor specific antibodies that are capable of inducing antibody-dependent cell-mediated cytotoxicity to allogeneic, as well as autologous, tumor cells. This ...
    DNA PCR Assays for Igh Rearrangement
    Authors:  Yusuke Satoh, Takao Sudo and Takafumi Yokota, date: 02/20/2014, view: 8461, Q&A: 0
    [Abstract] This protocol is used for the detection of immunoglobulin heavy (H) chain rearrangements. This PCR-based assay enables detection of DH-JH recombination in cultured hematopoietic cells (Schlissel et al., 1991; Satoh et al., 2013) [e.g. ES-derived cells (Satoh et al., 2013)].

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