Protocols in Current Issue
    In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor
    Authors:  Taisho Yamada and Akinori Takaoka, date: 01/05/2017, view: 5058, Q&A: 0
    [Abstract] Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR ...
    Assessment of Brown Adipocyte Thermogenic Function by High-throughput Respirometry
    Authors:  Kiana Mahdaviani, Ilan Benador and Orian Shirihai, date: 11/05/2015, view: 10137, Q&A: 0
    [Abstract] Brown adipose tissue (BAT) has the unique ability to dramatically increase mitochondrial uncoupled fuel oxidation for thermogenesis in response to adrenergic stimulation. A key parameter in assessing brown adipocyte thermogenic capacity is mitochondrial uncoupling as determined by respiration. Measuring mitochondrial oxygen consumption rate (OCR) ...
    Bone Marrow Derived Eosinophil Cultures
    Authors:  Thomas X. Lu and Marc E. Rothenberg, date: 06/20/2014, view: 10021, Q&A: 1
    [Abstract] Eosinophils are multifunctional effector cells implicated in the pathogenesis of a variety of diseases including asthma, eosinophil gastrointestinal disorders and helminth infection. Mouse bone marrow derived progenitor cells can be differentiated into eosinophils following IL-5 exposure. These bone marrow derived eosinophils are fully ...
    Small-scale Subcellular Fractionation with Sucrose Step Gradient
    Authors:  Yuzuru Taguchi and Hermann M. Schätzl, date: 06/05/2014, view: 12410, Q&A: 0
    [Abstract] Here, we introduce the protocol for small-scale and simple subcellular fractionation used in our recent publication (Taguchi et al., 2013), which uses homogenization by passing through needles and sucrose step-gradient.

    Subcellular fractionation is a very useful technique but usually a large number of cells are required. ...
    Isolation and Immortalization of Fibroblasts from Different Tumoral Stages
    Authors:  Fernando Calvo, Steven Hooper and Erik Sahai, date: 04/05/2014, view: 13804, Q&A: 0
    [Abstract] Tumour microenvironment and cancer-associated fibroblasts in particular exhibit tumour promoting abilities that are not present in their normal counterparts (Calvo et al., 2013; Hanahan and Coussens, 2012). Therefore, functional and molecular characterization of the modifications occurring in fibroblasts during tumour progression is ...
    Alkaline Phosphatase Staining
    Author:  Pearl A. Campbell, date: 03/05/2014, view: 22689, Q&A: 0
    [Abstract] Two main features characterize pluripotent cells; self-renewal (unlimited cell division) and the ability to give rise to all cells of the adult organism. Given the recent impact of induced pluripotent stem cells (iPSCs) and ongoing use of pluripotent embryonic stem cells ESCs (ESCs) in basic discovery, drug development, and potential use for stem ...
    Isolation of Mammary Epithelial Cells and Fibroblasts From Mouse Tumor
    Author:  Shiva Kazerounian, date: 02/20/2014, view: 15655, Q&A: 0
    [Abstract] Developing cancer therapeutics requires the ability to investigate their effects using in vitro models of a specific type of tumor. This protocol provides a method for the isolation and adoption to growth in culture of cells from primary tumors. This is particularly valuable for studying mouse models where original tumor cells can be ...
    Mitochondrial Transmembrane Potential (ψm) Assay Using TMRM
    Authors:  Isaline Rowe and Alessandra Boletta, date: 12/05/2013, view: 22057, Q&A: 0
    [Abstract] During cellular respiration, nutrients are oxidized to generate energy through a mechanism called oxidative phosphorylation, which occurs in the mitochondria. During oxidative phosphorylation, the gradual degradation of molecules through the TCA cycle releases electrons from the covalent bonds that are broken. These electrons are captured by NAD+ ...
    Isolation of Mouse Tumor-Infiltrating Leukocytes by Percoll Gradient Centrifugation
    Authors:  Ying Liu, Keqiang Chen, Chunyan Wang, Wanghua Gong, Teizo Yoshimura, Ji Ming Wang and Mingyong Liu, date: 09/05/2013, view: 32515, Q&A: 0
    [Abstract] A hallmark of cancer-associated inflammation is the infiltration of leukocytes into tumors, which is believed to be recruited by chemokines. Some infiltrating leukocytes such as macrophages often promote tumor growth by producing growth-inducing and angiogenic factors. Here, we described a method of isolating tumor-infiltrating leukocytes with ...
    Natural Killer Cell Transfer Assay
    [Abstract] Natural Killer (NK) cells are cytotoxic lymphocytes that constitute a major component of the innate immune system. Immunosurveillance of the host by NK cells for malignant and virally-infected cells results in direct cytotoxicity and the production of cytokines to enhance the immune response. This protocol will describe the adoptive transfer of ...

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