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    Protocols in Current Issue
    Chromatographic Assays for the Enzymatic Degradation of Chitin
    Authors:  Sophanit Mekasha, Tina R. Tuveng, Gustav Vaaje-Kolstad and Vincent G. H. Eijsink, date: 05/05/2021, view: 512, Q&A: 0
    [Abstract]

    Chitin is an insoluble linear polymer of β(1→4)-linked N-acetylglucosamine. Enzymatic cleavage of chitin chains can be achieved using hydrolytic enzymes, called chitinases, and/or oxidative enzymes, called lytic polysaccharide monooxygenases (LPMOs). These two groups of enzymes have different modes of action and yield different product

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    AIMTOR, a BRET Biosensor for Live Recording of mTOR Activity in Cell Populations and Single Cells
    Authors:  Nathalie Bouquier, Enora Moutin, Julie Perroy and Vincent Ollendorff, date: 04/20/2021, view: 913, Q&A: 0
    [Abstract]

    Mammalian target of rapamycin (mTOR) controls many crucial cellular functions, including protein synthesis, cell size, energy metabolism, lysosome and mitochondria biogenesis, and autophagy. Consequently, deregulation of mTOR signaling plays a role in numerous pathological conditions such as cancer, metabolic disorders and neurological diseases.

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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: 1021, 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

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    K+ Release Assay and K+ Measurement in Oocyte Assay
    Author:  Hong Li, date: 11/05/2020, view: 1143, Q&A: 0
    [Abstract] The Xenopus oocyte is a powerful system for the exogenous expression and functional characterization of plant membrane transport proteins. Until now, a number of potassium transporters and channels have been identified in oocytes expression system by the two-electrode voltage clamp technology. It is difficult to characterize K+ ...
    Probe-Seq: Method for RNA Sequencing of Specific Cell Types from Animal Tissue
    [Abstract] Most organs and tissues are composed of many types of cells. To characterize cellular state, various transcription profiling approaches are currently available, including whole-tissue bulk RNA sequencing, single cell RNA sequencing (scRNA-Seq), and cell type-specific RNA sequencing. What is missing in this repertoire is a simple, versatile method ...
    Long-term Devocalization of Zebra Finches
    Authors:  Yunbok Kim, Daisuke Mizuguchi and Satoshi Kojima, date: 09/20/2020, view: 1725, Q&A: 0
    [Abstract] Songbirds, such as the zebra finch, are a popular animal model for studying the neural basis of vocal and complex skill learning. Adult male zebra finches produce courtship song toward females (referred to as ‘directed song’) and recording and analyzing sounds of directed song along with underlying neural activity is important for investigating ...
    Minimally Invasive Oral Surgery Induction of the FRICT-ION Chronic Neuropathic Pain Model
    Authors:  Marena A. Montera and Karin N. Westlund, date: 04/20/2020, view: 1820, Q&A: 0
    [Abstract] An easily induced preclinical trigeminal neuropathic nerve injury model is described here for the study of chronic pain, the model acronym FRICT-ION (Foramen Rotundum Inflammatory Constriction Trigeminal InfraOrbital Nerve). In patients, neuropathic pain is thought to be related to ...
    Fractionation of Ovarian Follicles and in vitro Oocyte Maturation and Ovulation Assay of Ciona intestinalis Type A
    Authors:  Shin Matsubara, Akira Shiraishi, Tomohiro Osugi, Tsuyoshi Kawada and Honoo Satake, date: 04/05/2020, view: 1953, Q&A: 0
    [Abstract] Ascidians are the closest living relatives of vertebrates (Delsuc et al., 2006; Satoh et al., 2014) and are important for the evolutionary study of the ovarian follicle development including oocyte maturation and ovulation. However, neither the endogenous factors nor the molecular mechanisms underlying the oocyte maturation and ...
    The Chick Chorioallantoic Membrane (CAM) Assay as a Three-dimensional Model to Study Autophagy in Cancer Cells
    Authors:  Félice A. Janser, Patricia Ney, Marta Teixeira Pinto, Rupert Langer and Mario P. Tschan, date: 07/05/2019, view: 5049, Q&A: 0
    [Abstract] The chick chorioallantoic membrane (CAM) is an extra-embryonic organ and thus well accessible for seeding and harvesting 3D cell cultures. Samples from CAM assays are suitable for protein and gene expression analysis as well as for immuno-histochemical studies. Here we present the CAM assay as a possible model to study autophagy in different types ...
    Microplastic Extraction from Marine Vertebrate Digestive Tracts, Regurgitates and Scats: A Protocol for Researchers from All Experience Levels
    Authors:  Amy L. Lusher and Gema Hernandez-Milian, date: 11/20/2018, view: 6851, Q&A: 0
    [Abstract] It is essential to provide a protocol for the separation and identification of microplastics in marine vertebrates (mammals, birds, turtles and fish) that is easy to follow and adaptable depending on research infrastructure. Digesting organic material is an effective way to analyze samples for microplastics. Presented here is an optimized protocol ...



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