Protocols in Current Issue
    TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4+ T Cells
    Authors:  Anwesha Sanyal, Vatsala S. Rangachar and Phalguni Gupta, date: 05/20/2019, view: 456, Q&A: 0
    [Abstract] The latent HIV-1 viral reservoir in resting CD4+ (rCD4+) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the ...
    Isolation and Culture of Mouse Lung ILC2s
    Authors:  Pauline Schmitt, Anais Duval, Emilie Mirey, Jean-Philippe Girard and Corinne Cayrol, date: 10/05/2018, view: 1660, Q&A: 0
    [Abstract] Group 2 Innate Lymphoid Cells (ILC2) play an important role in immune responses at barrier surfaces, notably in the lung during airway allergic inflammation or asthma. Several studies have described methods to isolate ILC2s from wild-type naive mice, most of them using cell sorting to obtain a pure population. Here, we describe in detail, a ...
    Using Stable Isotopes in Bone Marrow Derived Macrophage to Analyze Metabolism
    [Abstract] Using gas chromatography mass spectrometry (GC-MS) to analyze the citric acid cycle (CAC) and related intermediates (such as glutamate, glutamine, GABA, and aspartate) is an analytical approach to identify unexpected correlations between apparently related and unrelated pathways of energy metabolism. Intermediates can be as expressed as their ...
    Phagocytosis Assay of Necroptotic Cells by Cardiac Myofibroblasts
    Authors:  Yuma Horii, Shoichi Matsuda, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose and Michio Nakaya, date: 09/20/2017, view: 3326, Q&A: 0
    [Abstract] In myocardial infarction (MI), a plenty of cardiomyocytes undergo necrosis and necroptosis due to the lack of oxygen and nutrients. The dead cardiomyocytes are promptly engulfed by phagocytes. When the dead cells are not engulfed, the noxious contents of the cells are released outside, and thus, induce inflammation, and obstruct the function of ...
    An Assay to Determine Phagocytosis of Apoptotic Cells by Cardiac Macrophages and Cardiac Myofibroblasts
    Authors:  Yuma Horii, Shoichi Matsuda, Kenji Watari, Akiomi Nagasaka, Hitoshi Kurose and Michio Nakaya, date: 09/20/2017, view: 3670, Q&A: 0
    [Abstract] In myocardial infarction (MI), a number of cardiomyocytes undergo apoptosis. These apoptotic cardiomyocytes are promptly engulfed by phagocytes. If the dead cells are not engulfed, their noxious contents are released outside, resulting in induction of inflammation. Therefore, the removal of these dead cells is necessary. However, the contribution ...
    Isolation and Analysis of Stromal Vascular Cells from Visceral Adipose Tissue
    Authors:  Jessica Vu and Wei Ying, date: 08/20/2017, view: 5767, Q&A: 0
    [Abstract] The obesity epidemic is the underlying driver of the type 2 diabetes mellitus epidemic. A remarkable accumulation of various pro-inflammatory immune cells in adipose tissues is a hallmark of obesity and leads to pathogenesis of tissue inflammation and insulin resistance. Here, we describe a detailed protocol to isolate adipose tissue stromal ...
    Isolation of PBMCs Using Vacutainer® Cellular Preparation Tubes (CPTTM)
    Authors:  Alaina Puleo, Chantia Carroll, Holden Maecker and Rohit Gupta, date: 01/20/2017, view: 16681, Q&A: 0
    [Abstract] Peripheral blood mononuclear cell (PBMC) isolation is commonly done via density gradient centrifugation over Ficoll-Hypaque, a labor-intensive procedure that requires skilled technicians and can contribute to sample variability. Cellular Preparation Tubes (CPTs) are Vacutainer blood draw tubes that contain Ficoll-Hypaque and a gel plug that ...
    Isolation of Highly Pure Primary Mouse Alveolar Epithelial Type II Cells by Flow Cytometric Cell Sorting
    Authors:  Meenal Sinha and Clifford A. Lowell, date: 11/20/2016, view: 11741, Q&A: 0
    [Abstract] In this protocol, we describe the method for isolating highly pure primary alveolar epithelial type II (ATII) cells from lungs of naïve mice. The method combines negative selection for a variety of lineage markers along with positive selection for EpCAM, a pan-epithelial cell marker. This method yields 2-3 x 106 ATII cells per mouse ...
    Isolation of Intestinal Mesenchymal Cells from Adult Mice
    Authors:  Vasiliki Koliaraki and George Kollias, date: 09/20/2016, view: 10003, Q&A: 0
    [Abstract] During the last 20 years intestinal mesenchymal cells (IMCs) have emerged as an important cell type that plays a central role in intestinal development and homeostasis, by providing both structural support and growth regulatory elements. IMCs also actively participate in wound healing responses, thus regulating pathologic conditions such as tissue ...
    Isolation of Joint-infiltrating Cells
    Authors:  Aoi Akitsu and Yoichiro Iwakura, date: 09/05/2016, view: 5402, Q&A: 0
    [Abstract] Infiltration of leukocytes into joints is one of the main features of autoimmune inflammatory arthritis. Here, we describe the protocol for isolation of joint-infiltrating cells in mice. This protocol is useful to analyze cell surface antigens and intracellular cytokines by flow cytometry.

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