Protocols in Current Issue
    Environmental Conditioning and Aerosol Infection of Mice
    Authors:  Eriko Kudo and Akiko Iwasaki, date: 04/20/2020, view: 1156, Q&A: 0
    [Abstract] Influenza infection models in mice are widely used to study flu-mediated immune responses and pathology. However, most laboratory mice are housed at 20 °C and 50% relative humidity (RH). To better recapitulate influenza epidemics and immune responses during winter seasons, mice were housed at 20 °C under different humidity conditions, 10-20% or ...
    A Triple-challenge Mouse Model of Allergic Airway Disease, Primary Influenza Infection, and Secondary Bacterial Infection
    Authors:  Sean Roberts, Clare M Williams, Sreeja Roy and Yoichi Furuya, date: 04/20/2020, view: 857, Q&A: 0
    [Abstract] Asthma is a global problem that affects millions of individuals. An increased risk of respiratory viral and bacterial infections is one of the complications of asthma. We recently reported that mice with ovalbumin-induced allergic airway disease (AAD) are protected against influenza-Streptococcus pneumoniae co-infection. Here, we describe ...
    Photoactivable Cholesterol as a Tool to Study Interaction of Influenza Virus Hemagglutinin with Cholesterol
    Authors:  Bodan Hu, Mohamed Rasheed Gadalla, Christoph Thiele and Michael Veit, date: 02/20/2020, view: 1385, Q&A: 0
    [Abstract] Non-covalent binding of cholesterol to the transmembrane region of proteins affect their functionalities, but methods to prove such an interaction are rare. We describe our protocol to label the hemagglutinin (HA) of Influenza virus with a cholesterol derivative in living cells or with immunoprecipitated protein. We synthesized a “clickable” ...
    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: 2163, 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: 2774, 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: 7180, 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: 7071, 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: 7201, 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 ...

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