Mammalia

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    Protocols in Current Issue
    SarkoSpin: A Technique for Biochemical Isolation and Characterization of Pathological TDP-43 Aggregates
    Authors:  Pérez-Berlanga Manuela, Laferrière Florent and Polymenidou Magdalini, date: 11/20/2019, view: 156, Q&A: 0
    TDP-43 is the main aggregating protein in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Aggregated TDP-43 is resistant to diverse detergent solubilization, yet physiological ...
    ELISA Based Protein Ubiquitylation Measurement
    Authors:  Yuka Kamada, Ryosuke Fukuda and Tsukasa Okiyoneda, date: 11/20/2019, view: 21, Q&A: 0
    Ubiquitylation is a common post-translational modification of cellular proteins that results in proteasomal and lysosomal degradations. Ubiquitylation is generally measured by methods such as immunoblotting using anti-ubiquitin antibodies after ...
    Bacterial Synchronized Transfer Assays in Bone Marrow Derived Macrophages
    Authors:  Sedelia R Dominguez, Tom Kawula and Shaun P Steele, date: 11/20/2019, view: 40, Q&A: 0
    Merocytophagy (“mero”, Greek for partial; “cytophagy” for cell eating) is a process by which cells acquire microbes and cytosolic material through phagocytosis of a small portion of neighboring cells upon cell-cell contact. Cell-cell contact ...
    Labeling and Isolation of Fluorouracil Tagged RNA by Cytosine Deaminase Expression
    Authors:  Harihar Basnet and Joan Massague, date: 11/20/2019, view: 25, Q&A: 0
    Tissues are comprised of different cell types whose interactions elicit distinct gene expression patterns that regulate tissue formation, regeneration, homeostasis and repair. Analysis of these gene expression patterns require methods that can ...
    Opto-magnetic Selection and Isolation of Single Cells
    Authors:  Loïc Binan, Joannie Roy and Santiago Costantino, date: 11/20/2019, view: 27, Q&A: 0
    Capturing single cells from large heterogenous populations based solely on observable traits is necessary for many cell biology applications and remains a major technical challenge. The protocol we present allows the isolation of viable and ...
    Isolation and Quantification of Metabolite Levels in Murine Tumor Interstitial Fluid by LC/MS
    Authors:  Mark R Sullivan, Caroline A Lewis and Alexander Muir, date: 11/20/2019, view: 132, Q&A: 0
    Cancer is a disease characterized by altered metabolism, and there has been renewed interest in understanding the metabolism of tumors. Even though nutrient availability is a critical determinant of tumor metabolism, there has been little systematic ...
    Leishmania Parasite Quantification by Bioluminescence in Murine Models
    Authors:  Marta Reverte and Nicolas Fasel, date: 11/20/2019, view: 29, Q&A: 0
    Leishmaniasis remains a major public health problem worldwide with a prevalence of 12 million, an incidence of 1 million persons, and 350 million people being at risk. Murine models have been largely used for studying the host-pathogen relationship ...
    SarkoSpin: A Technique for Biochemical Isolation and Characterization of Pathological TDP-43 Aggregates
    Authors:  Pérez-Berlanga Manuela, Laferrière Florent and Polymenidou Magdalini, date: 11/20/2019, view: 156, Q&A: 0
    [Abstract] TDP-43 is the main aggregating protein in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Aggregated TDP-43 is resistant to diverse detergent solubilization, yet physiological TDP-43 and other abundant proteins commonly co-purify with pathological TDP-43. This mixed isolation has ...
    ELISA Based Protein Ubiquitylation Measurement
    Authors:  Yuka Kamada, Ryosuke Fukuda and Tsukasa Okiyoneda, date: 11/20/2019, view: 21, Q&A: 0
    [Abstract] Ubiquitylation is a common post-translational modification of cellular proteins that results in proteasomal and lysosomal degradations. Ubiquitylation is generally measured by methods such as immunoblotting using anti-ubiquitin antibodies after isolating the protein-of-interest by denaturing immunoprecipitation. The following protocol can be used ...
    Bacterial Synchronized Transfer Assays in Bone Marrow Derived Macrophages
    Authors:  Sedelia R Dominguez, Tom Kawula and Shaun P Steele, date: 11/20/2019, view: 40, Q&A: 0
    [Abstract] Merocytophagy (“mero”, Greek for partial; “cytophagy” for cell eating) is a process by which cells acquire microbes and cytosolic material through phagocytosis of a small portion of neighboring cells upon cell-cell contact. Cell-cell contact dependent transfer events can be assessed through co-incubation of differently labeled cells. With these ...
    Labeling and Isolation of Fluorouracil Tagged RNA by Cytosine Deaminase Expression
    Authors:  Harihar Basnet and Joan Massague, date: 11/20/2019, view: 25, Q&A: 0
    [Abstract] Tissues are comprised of different cell types whose interactions elicit distinct gene expression patterns that regulate tissue formation, regeneration, homeostasis and repair. Analysis of these gene expression patterns require methods that can capture as closely as possible the transcriptomes of cells of interest in their tissue microenvironment. ...
    Opto-magnetic Selection and Isolation of Single Cells
    Authors:  Loïc Binan, Joannie Roy and Santiago Costantino, date: 11/20/2019, view: 27, Q&A: 0
    [Abstract] Capturing single cells from large heterogenous populations based solely on observable traits is necessary for many cell biology applications and remains a major technical challenge. The protocol we present allows the isolation of viable and metabolically active cells selected for their shape, migration speed, contact to other cells, or ...
    Isolation and Quantification of Metabolite Levels in Murine Tumor Interstitial Fluid by LC/MS
    Authors:  Mark R Sullivan, Caroline A Lewis and Alexander Muir, date: 11/20/2019, view: 132, Q&A: 0
    [Abstract] Cancer is a disease characterized by altered metabolism, and there has been renewed interest in understanding the metabolism of tumors. Even though nutrient availability is a critical determinant of tumor metabolism, there has been little systematic study of the nutrients directly available to cancer cells in the tumor microenvironment. Previous ...
    Leishmania Parasite Quantification by Bioluminescence in Murine Models
    Authors:  Marta Reverte and Nicolas Fasel, date: 11/20/2019, view: 29, Q&A: 0
    [Abstract] Leishmaniasis remains a major public health problem worldwide with a prevalence of 12 million, an incidence of 1 million persons, and 350 million people being at risk. Murine models have been largely used for studying the host-pathogen relationship and developing effective chemotherapies against Leishmania parasites. Thus, preclinical ...
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 308, Q&A: 0
    [Abstract] Hepatic stellate cells (HSCs), alternatively known as liver pericytes, can differentiate into myofibroblasts and secrete extra-cellular matrix components, thereby promoting wound healing and fibrosis. Studying HSCs can provide insights into the pathological mechanisms governing these processes. HSC isolation methods typically comprise of enzymatic ...
    Adoptive Transfer of Basophils Enriched from Mouse Spleen
    Authors:  Adrian M. Piliponsky and Stephen J. Galli, date: 11/05/2019, view: 138, Q&A: 0
    [Abstract] CD49b is a member of the integrin family, expressed on basophils, natural killer (NK) cells and a subset CD4+ T cells in the spleen. This protocol describes the adoptive transfer of basophil-enriched CD49b+ cells obtained from mouse spleens by magnetic enrichment. This protocol can be used to assess the contribution of ...
    Imaging VIPER-labeled Cellular Proteins by Correlative Light and Electron Microscopy
    [Abstract] Advances in fluorescence microscopy (FM), electron microscopy (EM), and correlative light and EM (CLEM) offer unprecedented opportunities for studying diverse proteins and nanostructures involved in fundamental cell biology. It is now possible to visualize and quantify the spatial organization of cellular proteins and other macromolecules by FM, ...



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