Cancer Biology

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    Protocols in Current Issue
    A Transient Transfection-based Cell Adhesion Assay with 293T Cells
    Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 464, Q&A: 0

    The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a

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    CENP-C Phosphorylation by CDK1 in vitro
    Authors:  Reito Watanabe, Masatoshi Hara, Mariko Ariyoshi and Tatsuo Fukagawa, date: 01/05/2021, view: 307, Q&A: 0

    Accurate chromosome segregation during mitosis requires the kinetochore, a large protein complex, which makes a linkage between chromosomes and spindle microtubes. An essential kinetochore component, CENP-C, is phosphorylated by Cyclin-B-Cyclin

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    A Rigorous Quantitative Approach to Analyzing Phagocytosis Assays
    Authors:  Michael D. Caponegro, Kaitlyn Koenig Thompson, Maryam Tayyab and Stella E. Tsirka, date: 01/05/2020, view: 1279, Q&A: 0
    Studying monocytic cells in isolated systems in vitro contributes significantly to the understanding of innate immune physiology. Functional assays produce read outs which can be used to measure responses to selected stimuli, such as ...
    CENP-C Phosphorylation by CDK1 in vitro
    Authors:  Reito Watanabe, Masatoshi Hara, Mariko Ariyoshi and Tatsuo Fukagawa, date: 01/05/2021, view: 307, Q&A: 0
    [Abstract]

    Accurate chromosome segregation during mitosis requires the kinetochore, a large protein complex, which makes a linkage between chromosomes and spindle microtubes. An essential kinetochore component, CENP-C, is phosphorylated by Cyclin-B-Cyclin dependent kinase 1 (CDK1) that is a master kinase for mitotic progression, promoting proper kinetochore

    ...
    A Transient Transfection-based Cell Adhesion Assay with 293T Cells
    Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 464, Q&A: 0
    [Abstract]

    The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a transient 293T cell transfection-based cell adhesion assay to study selective cell adhesion of a

    ...
    Equilibrium and Kinetic Measurements of Ligand Binding to HiBiT-tagged GPCRs on the Surface of Living Cells
    Authors:  Michelle E. Boursier, Sergiy Levin, Robin Hurst and Rachel Friedman Ohana, date: 12/20/2020, view: 540, Q&A: 0
    [Abstract]

    G-protein coupled receptors (GPCRs) remain at the forefront of drug discovery efforts. Detailed assessment of features contributing to GPCR ligand engagement in a physiologically relevant environment is imperative to the development of new therapeutics with improved efficacy. Traditionally, binding properties such as affinity and kinetics were

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    Triacylglycerol Measurement in HeLa Cells
    Authors:  Ximing Du and Hongyuan Yang, date: 12/20/2020, view: 715, Q&A: 0
    [Abstract]

    Lipid droplets store triacylglycerols (triglycerides) and sterol esters to regulate lipid and energy homeostasis. Triacylglycerol measurement is often performed during the investigation of lipid droplet formation and growth. This protocol describes a reliable method using a fluorometric lipid quantification kit to measure triacylglycerols

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    A 3D Skin Melanoma Spheroid-Based Model to Assess Tumor-Immune Cell Interactions
    Authors:  Marek Wagner and Shigeo Koyasu, date: 12/05/2020, view: 709, Q&A: 0
    [Abstract] Three-dimensional (3D) tumor spheroids have the potential to bridge the gap between two-dimensional (2D) monolayer tumor cell cultures and solid tumors with which they share a significant degree of similarity. However, the progression of solid tumors is often influenced by the dynamic and reciprocal interactions between tumor and immune cells. ...
    Magnet-assisted Flow Cytometry of in vivo Tumors to Quantitate Cell-specific Responses to Magnetic Iron Oxide Nanoparticles
    Authors:  Preethi Korangath and Robert Ivkov, date: 11/20/2020, view: 664, Q&A: 0
    [Abstract] A clear understanding of nanoparticle interactions with living systems at the cellular level is necessary for developing nanoparticle-based therapeutics. Magnetic iron oxide nanoparticles provide unique opportunities to study these interactions because of their responsiveness to magnetic fields. This enables sorting of cells containing ...
    Dissecting the Rat Mammary Gland: Isolation, Characterization, and Culture of Purified Mammary Epithelial Cells and Fibroblasts
    [Abstract]

    With the advent of CRISPR-Cas and the ability to easily modify the genome of diverse organisms, rat models are being increasingly developed to interrogate the genetic events underlying mammary development and tumorigenesis. Protocols for the isolation and characterization of mammary epithelial cell subpopulations have been thoroughly developed for

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    Advances in Proximity Ligation in situ Hybridization (PLISH)
    Authors:  Monica Nagendran, Adam M Andruska, Pehr B Harbury and Tushar J Desai, date: 11/05/2020, view: 993, Q&A: 0
    [Abstract] Understanding tissues in the context of development, maintenance and disease requires determining the molecular profiles of individual cells within their native in vivo spatial context. We developed a Proximity Ligation in situ Hybridization technology (PLISH) that enables quantitative measurement of single cell gene expression ...
    Real-time Three-dimensional Tracking of Endocytic Vesicles
    Authors:  Stephanie Duhamel and Kossay Zaoui, date: 10/20/2020, view: 711, Q&A: 0
    [Abstract] Endocytic trafficking and recycling are fundamental cellular processes that control essential functions such as signaling protein complexes transport and membrane identity. The small GTPase Rabs are indispensable component of the endosomal recycling machinery. The Rabs bind to effectors to mediate their functions, such as protein sorting and ...
    Quantification of Salivary Charged Metabolites Using Capillary Electrophoresis Time-of-flight-mass Spectrometry
    Authors:  Masahiro Sugimoto, Sana Ota, Miku Kaneko, Ayame Enomoto and Tomoyoshi Soga, date: 10/20/2020, view: 610, Q&A: 0
    [Abstract] Salivary metabolomics have provided the potentials to detect both oral and systemic diseases. Capillary electrophoresis time-of-flight-mass spectrometry (CE-TOFMS) enables the identification and quantification of various charged metabolites. This method has been employed to biomarker discoveries using human saliva samples, especially for various ...



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