Protocols in Current Issue
    Evaluation of B Cell Proliferation in vivo by EdU Incorporation Assay
    Authors:  Adi Biram and Ziv Shulman, date: 05/05/2020, view: 784, Q&A: 0
    [Abstract] Generation of antibodies is crucial for establishing enduring protection from invading pathogens, as well as for maintaining homeostasis with commensal bacteria at mucosal surfaces. Chronic exposure to microbiota- and dietary- derived antigens results in continuous production of antibody producing cells within the Peyer’s patch germinal center ...
    Isolation and Characterization of Exosomes from Mouse Feces
    Authors:  Chunhua Yang, Mingzhen Zhang, Junsik Sung, Lixin Wang, Yunjin Jung and Didier Merlin, date: 04/20/2020, view: 921, Q&A: 0
    [Abstract] Exosomes secreted by colonic epithelial cells are present in feces and contain valuable epigenetic information, such as miRNAs, proteins, and metabolites. An in-depth study of this information is conducive to the diagnosis or treatment of relevant diseases. A crucial prerequisite of such a study is to establish an efficient isolation method, ...
    Characterization of Immunological Niches within Peyer’s Patches by ex vivo Photoactivation and Flow Cytometry Analysis
    Authors:  Adi Biram and Ziv Shulman, date: 03/20/2020, view: 701, Q&A: 0
    [Abstract] T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and involves physical contacts between T and B cells. During these interactions, T cells endow the B cells ...
    In vitro Self-organized Mouse Small Intestinal Epithelial Monolayer Protocol
    Authors:  Gizem Altay, Eduard Batlle, Vanesa Fernández-Majada and Elena Martinez, date: 02/05/2020, view: 2005, Q&A: 0
    [Abstract] Developing protocols to obtain intestinal epithelial monolayers that recapitulate in vivo physiology to overcome the limitations of the organoids’ closed geometry has become of great interest during the last few years. Most of the developed culture models showed physiological-relevant cell composition but did not prove self-renewing ...
    Induction and Analysis of Anti-CD40-induced Colitis in Mice
    Authors:  Barbara Joyce-Shaikh, Daniel J. Cua and David Bauché, date: 02/05/2019, view: 3810, Q&A: 0
    [Abstract] Colon inflammation or colitis affects more than 1 million people worldwide. Several pre-clinical models, including chemical-induced (i.e., DSS, TNBS) or pathogen-induced (i.e., Citrobacter rodentium) have been used to study mechanisms involved in the development and regulation of colitis. Anti-CD40 induced colitis model ...
    Use of Gas Chromatography to Quantify Short Chain Fatty Acids in the Serum, Colonic Luminal Content and Feces of Mice
    [Abstract] Short-Chain Fatty Acids (SCFAs) are a product of the fermentation of resistant starches and dietary fibers by the gut microbiota. The most important SCFA are acetate (C2), propionate (C3) and butyrate (C4). These metabolites are formed and absorbed in the colon and then transported through the hepatic vein to the liver. SCFAs are more concentrated ...
    Eicosanoid Isolation from Mouse Intestinal Tissue for ELISA
    Author:  Isabella Rauch, date: 11/05/2018, view: 2007, Q&A: 0
    [Abstract] Activation of inflammasomes in peritoneal macrophages and intestinal epithelial cells (IEC) leads to the release of eicosanoids. To assess the amount of eicosanoids released by IEC, lipids need to be isolated from whole tissue previous to analysis by lipid mass spectrometry or ELISA. This protocol describes how to isolate lipids from intestinal ...
    Purification of Total RNA from DSS-treated Murine Tissue via Lithium Chloride Precipitation
    Authors:  Emilie Viennois, Anika Tahsin and Didier Merlin, date: 05/05/2018, view: 4205, Q&A: 0
    [Abstract] We have developed a protocol to purify RNA from DSS (Dextran Sulfate Sodium)-treated mouse tissues. This method, which prevents downstream inhibition of q-RT-PCR observed in DSS-treated tissues, relies on successive precipitations with lithium chloride.
    Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection
    Authors:  Jana Van Dycke, Johan Neyts and Joana Rocha-Pereira, date: 05/05/2018, view: 3396, Q&A: 0
    [Abstract] Human norovirus is the most common cause of acute gastroenteritis worldwide, resulting in estimated mortality of ~210,000 each year, of whom most are children under the age of five. However, norovirus can infect people of all age groups. There is a risk of prolonged infection in children, the elderly and patients who are immunocompromised. To ...
    Quantification of Bacterial Attachment to Tissue Sections
    Authors:  Batya Isaacson, Tehila Hadad, Gilad Bachrach and Ofer Mandelboim, date: 03/05/2018, view: 4393, Q&A: 0
    [Abstract] Here we describe a method to test bacterial adhesion to paraffin embedded tissue sections. This method allows examining binding of different bacterial strains, transfected with a fluorescent protein reporter plasmid to various tissues, to better understand different mechanisms such as colonization. This assay provides a more physiological context ...

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