Mammalia

Categories

    Protocols in Current Issue
    Triple Fluorescence Anisotropy Reporter Imaging in Living Cells
    Authors:  Brian Ross, Shenq Huey Wong, Nicole E. Snell, Jin Zhang and M. A. Rizzo, date: 05/05/2019, view: 654, Q&A: 0
    [Abstract] FRET-based genetically encoded biosensors incorporate two fluorescent proteins into their design to enable ratiometric biosensing of signaling activities in live cells. While emission ratios are generally useful for quantitative studies, they leave little room in the optical spectrum for additional sensors and optogenetic tools. Homotransfer-based ...
    Structural Analysis of Target Protein by Substituted Cysteine Accessibility Method
    Authors:  Tetsuo Cai and Taisuke Tomita, date: 09/05/2018, view: 1456, Q&A: 0
    [Abstract] Substituted Cysteine Accessibility Method (SCAM) is a biochemical approach to investigate the water accessibility or the spatial distance of particular cysteine residues substituted in the target protein. Protein topology and structure can be annotated by labeling with methanethiosulfonate reagents that specifically react with the cysteine ...
    Measuring Mitochondrial ROS in Mammalian Cells with a Genetically Encoded Protein Sensor
    [Abstract] Reactive oxygen species (ROS) are not only known for their toxic effects on cells, but they also play an important role as second messengers. As such, they control a variety of cellular functions such as proliferation, metabolism, differentiation and apoptosis. Thus, ROS are involved in the regulation of multiple physiological and ...
    Protocol for HeLa Cells Infection with Escherichia coli Strains Producing Colibactin and Quantification of the Induced DNA-damage
    [Abstract] Strains of Escherichia coli bearing the pks genomic island synthesize the genotoxin colibactin. Exposure of eukaryotic cells to E. coli producing colibactin induces DNA damages, ultimately leading to cell cycle arrest, senescence and death. Here we describe a simple method to demonstrate the genotoxicity of bacteria ...
    Ca2+ Measurements in Mammalian Cells with Aequorin-based Probes
    Authors:  Anna Tosatto, Rosario Rizzuto and Cristina Mammucari, date: 03/05/2017, view: 4115, Q&A: 0
    [Abstract] Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca2+ uptake and release of different ...
    Assessment of Cellular Redox State Using NAD(P)H Fluorescence Intensity and Lifetime
    Authors:  Thomas S. Blacker, Tunde Berecz, Michael R. Duchen and Gyorgy Szabadkai, date: 01/20/2017, view: 6543, Q&A: 0
    [Abstract] NADH and NADPH are redox cofactors, primarily involved in catabolic and anabolic metabolic processes respectively. In addition, NADPH plays an important role in cellular antioxidant defence. In live cells and tissues, the intensity of their spectrally-identical autofluorescence, termed NAD(P)H, can be used to probe the mitochondrial redox state, ...
    Reconstruction of the Mouse Inflammasome System in HEK293T Cells
    Authors:  Hexin Shi, Anne Murray and Bruce Beutler, date: 11/05/2016, view: 5252, Q&A: 0
    [Abstract] The NLRP3 (NLR family, Pyrin domain containing 3) inflammasome is a multiprotein complex comprised of NLRP3, pro-caspase-1, the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), and the protein kinase NIMA related kinase 7 (NEK7) (Shi et al., 2016; He et al., 2016; Schmid-Burgk et al., ...
    Chromium-51 (51Cr) Release Assay to Assess Human T Cells for Functional Avidity and Tumor Cell Recognition
    Authors:  Petra Baumgaertner, Daniel E. Speiser, Pedro Romero, Nathalie Rufer and Michael Hebeisen, date: 08/20/2016, view: 9372, Q&A: 0
    [Abstract] Cytotoxic CD8+ T cells are able to specifically recognize and kill target cells through specific interaction between their T cell receptors (TCRs) and small immunogenic peptides (antigens) presented by major histocompatibility complex (MHC) molecules. The antigen recognition capacity and in vitro lytic activity of ...
    Enzymatic Activity Assays for Base Excision Repair Enzymes in Cell Extracts from Vertebrate Cells
    Authors:  Melike Çağlayan, Julie K. Horton and Samuel H. Wilson, date: 06/05/2015, view: 4909, Q&A: 0
    [Abstract] We previously reported enzymatic activity assays for the base excision repair (BER) enzymes DNA polymerase β (pol β), aprataxin (APTX), and flap endonuclease 1 (FEN1) in cell extracts from Saccharomyces cerevisiae (Çağlayan and Wilson, 2014). Here, we describe a method to prepare cell extracts from vertebrate cells to investigate these ...
    Glioma Associated Stem Cells (GASCs) Isolation and Culture
    [Abstract] Glioma Associated Stem Cells (GASCs) represent a population of non-tumorigenic multipotent stem cells hosted in the microenvironment of human gliomas. In vitro, these cells are able, through the release of exosomes, to increase the biological aggressiveness of glioma-initiating cells. The clinical importance of this finding is supported ...



    We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.