Protocols in Current Issue
    Production of Quantum Dots-containing Influenza Virus Particles for Studying Viral Uncoating Processes
    Authors:  Chong Qin, Xiaowei Zhang and Zongqiang Cui, date: 09/20/2019, view: 168, Q&A: 0
    [Abstract] The genome of influenza A virus (IAV) comprises eight pinlike genomic segments called vRNPs enclosed in viral capsid. During infection, uncoating is the key step for viral replication and represents an antiviral therapeutic target, but it is difficult to observe the transient and dynamic event in detail. Here, we report a protocol for production ...
    H1N1 Virus Production and Infection
    Authors:  Binbin Zhao, Jiaoyu Shan, Rui Xiong, Ke Xu and Bin Li, date: 10/20/2018, view: 1561, Q&A: 0
    [Abstract] Influenza A virus is a member of orthomyxoviridae family causing wide-spread infections in human respiratory tract. Mouse infection model is widely used in antiviral research and pathogenesis study against influenza A virus. Here, we report a protocol in infected mice with different virus doses and strains to explore how an inhibitor of ...
    Purification and Identification of Novel Host-derived Factors for Influenza Virus Replication from Human Nuclear Extracts
    Authors:  Kenji Sugiyama and Kyosuke Nagata, date: 09/20/2016, view: 5702, Q&A: 0
    [Abstract] Recently, we identified two host cell-derived proteins as novel stimulatory factors of influenza virus RNA replication process, termed “Influenza virus REplication Factor-2 (IREF-2)”, from human nuclear extracts (NEs) by employing biochemical complementation assays (Sugiyama et al., 2015). Herein, we describe detailed methods for ...
    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 ...
    A SYBR Green-based Real Time RT-PCR Assay for Detection of the Emerging H7N9 Virus
    Authors:  Zheng Zhu and Lunbiao Cui, date: 06/20/2014, view: 6000, Q&A: 0
    [Abstract] Most recently a novel avian-origin influenza A (H7N9) virus emerged in China and has been associated with lots of human infection and fatal cases. Molecular diagnostic methods are thus urgently needed in public health laboratories. We developed a SYBR green-based one-step real time reverse transcription-PCR (RT-PCR) to detect the novel H7N9 virus.
    Zonal Sedimentation Analysis on Sucrose Gradients
    Author:  Javier G. Magadán, date: 04/20/2014, view: 6389, Q&A: 0
    [Abstract] Zonal sedimentation analysis on sucrose gradients allows estimation of the molecular size of an individual protein or a protein complex by centrifugation at a constant speed under nondenaturing conditions. This method is particularly suitable for globular proteins like the influenza A virus (IAV) protein hemagglutinin (HA). Here, I describe step ...
    Radioactive Pulse-Chase Analysis and Immunoprecipitation
    Author:  Javier G. Magadán, date: 04/20/2014, view: 8909, Q&A: 0
    [Abstract] Labeling of newly-synthesized polypeptides with radioactive amino acids followed by immunoprecipitation allows quantitative analysis of the fate of a given protein in a time-dependent manner. This biochemical approach is usually used to study a variety of processes, such as protein folding, co-translational modifications, intracellular transport, ...
    ADCC Assay Protocol
    Authors:  Vikram Srivastava, Zheng Yang, Ivan Fan Ngai Hung, Jianqing Xu, Bojian Zheng and Mei-Yun Zhang, date: 01/20/2014, view: 25430, Q&A: 0
    [Abstract] Antibody-dependent cell-mediated cytotoxicity (ADCC) bridges innate and adaptive immunity, and it involves both humoral and cellular immune responses. ADCC has been found to be a main route of immune protection against viral infections and cancers in vivo. Here we developed a flow cytometry based protocol for ADCC assay using human ...
    Influenza Virus-cell Fusion Inhibition Assay
    Authors:  Mayo Yasugi and Kazuyoshi Ikuta, date: 12/20/2013, view: 7220, Q&A: 0
    [Abstract] During viral infection to host cells, several viruses undergo the process of endocytosis and pH-dependent fusion. By fusion of viral membrane with host cellular membrane, the viral core invades to host cytoplasm. A part of monoclonal antibodies against viral membrane protein have potential to inhibit the viral fusion step. Here we describe in ...

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