Search

Mammalia

Categories

    Protocols in Current Issue
    An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist
    [Abstract] One fundamental property of the TNR receptor (TNFR) family relates to how ‘signal quality’ (the extent of receptor ligation or cross-linking) influences the outcome of receptor ligation, for instance the induction of death in tumour cells. It is unequivocal that membrane-presented ligand (delivered to target cells via cell-surface presentation by ...
    Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA
    Authors:  Christian Reuter and Tobias A. Oelschlaeger, date: 06/20/2018, view: 449, Q&A: 0
    [Abstract] The mucosal surfaces of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye harbor a resident microflora that lives in symbiosis with their host and forms a complex ecosystem. The protection of the vulnerable epithelium is primarily achieved by mucins that form a gel-like structure adherent to the apical ...
    Intracellular and Mitochondrial Reactive Oxygen Species Measurement in Primary Cultured Neurons
    [Abstract] Reactive oxygen species (ROS) are chemically reactive oxygen containing molecules. ROS consist of radical oxygen species including superoxide anion (O2•−) and hydroxyl radical (•OH) and non-radical oxygen species such as hydrogen peroxide (H2O2), singlet oxygen (O2). ROS are generated by ...
    Expansion of Airway Basal Cells and Generation of Polarized Epithelium
    Authors:  Hannah Levardon, Lael M. Yonker, Bryan P. Hurley and Hongmei Mou, date: 06/05/2018, view: 368, Q&A: 0
    [Abstract] Airway basal stem cells are the progenitor cells within the airway that exhibit the capacity to self-renew and give rise to multiple types of differentiated airway epithelial cells. This stem cell-derived epithelium displays organized architecture with functional attributes of the airway mucosa. A protocol has been developed to culture and expand ...
    Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
    Authors:  Birte Plitzko and Sandra Loesgen, date: 05/20/2018, view: 1253, Q&A: 0
    [Abstract] Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual ...
    Mammalian Cell-derived Vesicles for the Isolation of Organelle Specific Transmembrane Proteins to Conduct Single Molecule Studies
    Authors:  Faruk H. Moonschi, Ashley M. Fox-Loe, Xu Fu and Chris I. Richards, date: 05/05/2018, view: 510, Q&A: 0
    [Abstract] Cell-derived vesicles facilitate the isolation of transmembrane proteins in their physiological membrane maintaining their structural and functional integrity. These vesicles can be generated from different cellular organelles producing, housing, or transporting the proteins. Combined with single-molecule imaging, isolated organelle specific ...
    An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells
    Authors:  Paula Szalai and Nikolai Engedal, date: 05/05/2018, view: 725, Q&A: 0
    [Abstract] In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. ...
    The Long-lived Protein Degradation Assay: an Efficient Method for Quantitative Determination of the Autophagic Flux of Endogenous Proteins in Adherent Cell Lines
    Authors:  Morten Luhr, Frank Sætre and Nikolai Engedal, date: 05/05/2018, view: 785, Q&A: 0
    [Abstract] Autophagy is a key player in the maintenance of cellular homeostasis in eukaryotes, and numerous diseases, including cancer and neurodegenerative disorders, are associated with alterations in autophagy. The interest for studying autophagy has grown intensely in the last two decades, and so has the arsenal of methods utilised to study this highly ...
    Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
    Authors:  Jorge Luis Ayala-Lujan and Fernando Ruiz-Perez, date: 04/20/2018, view: 574, Q&A: 0
    [Abstract] Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory ...
    A Method for Extracting the Nuclear Scaffold from the Chromatin Network
    Authors:  Junjie Chen, Boon Heng Dennis Teo and Jinhua Lu, date: 04/20/2018, view: 623, Q&A: 0
    [Abstract] Each cell contains many large DNA polymers packed in a nucleus of approx. 10 μm in diameter. With histones, these DNA polymers are known to form chromatins. How chromatins further compact in the nucleus is unclear but it inevitably depends on an extensive non-chromatin nuclear scaffold. Imaging of endogenous chromatin network and the complementary ...