Mammalia

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    Protocols in Current Issue
    Skin Transplantation and Lymphoid Organ Analysis in Mice
    Skin transplantation in mice is an important procedure to evaluate immune responses generated against heterologous grafts, especially given its highly immunogenic nature. In fact, skin is one of the most challenging organs in terms of allograft ...
    Estimating Cellular Abundances of Halo-tagged Proteins in Live Mammalian Cells by Flow Cytometry
    Authors:  Claudia Cattoglio, Xavier Darzacq, Robert Tjian and Anders Sejr Hansen, date: 02/20/2020, view: 258, Q&A: 0
    Accurate abundance measurements of cellular proteins are required to achieve a quantitative and predictive understanding of any biological process inside the cell. Existing methods to determine absolute protein abundances are labor-intensive and/or ...
    Assessing Self-interaction of Mammalian Nuclear Proteins by Co-immunoprecipitation
    Authors:  Claudia Cattoglio, Iryna Pustova, Xavier Darzacq, Robert Tjian and Anders S. Hansen, date: 02/20/2020, view: 236, Q&A: 0
    Protein-protein interactions constitute the molecular foundations of virtually all biological processes. Co-immunoprecipitation (CoIP) experiments are probably the most widely used method to probe both heterotypic and homotypic protein-protein ...
    Preparation of Single Epithelial Cells Suspension from Mouse Mammary Glands
    Authors:  Heng Sun, Xiaoling Xu and Chuxia Deng, date: 02/20/2020, view: 55, Q&A: 0
    Single cell RNA sequencing is a very powerful means for cellular heterogeneous studies and so becoming wildly utilized nowadays. To guarantee the success of such analysis, it is very important, though sometimes difficult, to obtain single cells ...
    Hippocampal Unicellular Recordings and Hippocampal-dependent Innate Behaviors in an Adolescent Mouse Model of Alzheimer’s disease
    Transgenic mice have been used to make valuable contributions to the field of neuroscience and model neurological diseases. The simultaneous functional analysis of hippocampal cell activity combined with hippocampal dependent innate task ...
    RNA Sequencing of Single Myofibers from Mus musculus
    Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 119, Q&A: 0
    Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but ...
    Induction of Temporal Lobe Epilepsy in Mice with Pilocarpine
    Authors:  Muhammad Nauman Arshad and Janice R Naegele, date: 02/20/2020, view: 51, Q&A: 0
    In the pilocarpine model of temporal lobe epilepsy (TLE) in rodents, systemic injections of pilocarpine induce continuous, prolonged limbic seizures, a condition termed “Status Epilepticus” (SE). With appropriate doses, many inbred strains of mice ...
    Method for Primary Epithelial Cell Culture from the Rat Choroid Plexus
    Authors:  Valeria Lallai, Amina Ahmed and Christie D. Fowler, date: 02/20/2020, view: 54, Q&A: 0
    The choroid plexus consists of a network of secretory epithelial cells localized throughout the lateral, third and fourth ventricles of the brain. Cerebrospinal fluid (CSF) is generated by the choroid plexus and released into the ventricular ...
    ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach
    Authors:  Junaid Akhtar, Piyush More and Steffen Albrecht, date: 02/20/2020, view: 98, Q&A: 0
    Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. ...
    Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation
    Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and ...
    Skin Transplantation and Lymphoid Organ Analysis in Mice
    [Abstract] Skin transplantation in mice is an important procedure to evaluate immune responses generated against heterologous grafts, especially given its highly immunogenic nature. In fact, skin is one of the most challenging organs in terms of allograft retention. In this protocol, we provide a detailed procedure for skin grafting using the tail skin as ...
    Estimating Cellular Abundances of Halo-tagged Proteins in Live Mammalian Cells by Flow Cytometry
    Authors:  Claudia Cattoglio, Xavier Darzacq, Robert Tjian and Anders Sejr Hansen, date: 02/20/2020, view: 258, Q&A: 0
    [Abstract] Accurate abundance measurements of cellular proteins are required to achieve a quantitative and predictive understanding of any biological process inside the cell. Existing methods to determine absolute protein abundances are labor-intensive and/or require sophisticated experimental and computational infrastructure (e.g., fluorescence ...
    Assessing Self-interaction of Mammalian Nuclear Proteins by Co-immunoprecipitation
    Authors:  Claudia Cattoglio, Iryna Pustova, Xavier Darzacq, Robert Tjian and Anders S. Hansen, date: 02/20/2020, view: 236, Q&A: 0
    [Abstract] Protein-protein interactions constitute the molecular foundations of virtually all biological processes. Co-immunoprecipitation (CoIP) experiments are probably the most widely used method to probe both heterotypic and homotypic protein-protein interactions. Recent advances in super-resolution microscopy have revealed that several nuclear proteins ...
    Preparation of Single Epithelial Cells Suspension from Mouse Mammary Glands
    Authors:  Heng Sun, Xiaoling Xu and Chuxia Deng, date: 02/20/2020, view: 55, Q&A: 0
    [Abstract] Single cell RNA sequencing is a very powerful means for cellular heterogeneous studies and so becoming wildly utilized nowadays. To guarantee the success of such analysis, it is very important, though sometimes difficult, to obtain single cells suspension with high quality, especially from the primary solid organs like mammary glands. Digestion of ...
    Hippocampal Unicellular Recordings and Hippocampal-dependent Innate Behaviors in an Adolescent Mouse Model of Alzheimer’s disease
    [Abstract] Transgenic mice have been used to make valuable contributions to the field of neuroscience and model neurological diseases. The simultaneous functional analysis of hippocampal cell activity combined with hippocampal dependent innate task evaluations provides a reliable experimental approach to detect fine changes during early phases of ...
    RNA Sequencing of Single Myofibers from Mus musculus
    Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 119, Q&A: 0
    [Abstract] Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the ...
    Induction of Temporal Lobe Epilepsy in Mice with Pilocarpine
    Authors:  Muhammad Nauman Arshad and Janice R Naegele, date: 02/20/2020, view: 51, Q&A: 0
    [Abstract] In the pilocarpine model of temporal lobe epilepsy (TLE) in rodents, systemic injections of pilocarpine induce continuous, prolonged limbic seizures, a condition termed “Status Epilepticus” (SE). With appropriate doses, many inbred strains of mice show behavioral seizures within an hour after pilocarpine is injected. With the behavioral scoring ...
    Method for Primary Epithelial Cell Culture from the Rat Choroid Plexus
    Authors:  Valeria Lallai, Amina Ahmed and Christie D. Fowler, date: 02/20/2020, view: 54, Q&A: 0
    [Abstract] The choroid plexus consists of a network of secretory epithelial cells localized throughout the lateral, third and fourth ventricles of the brain. Cerebrospinal fluid (CSF) is generated by the choroid plexus and released into the ventricular environment. This biofluid contains an enriched source of proteins, ions, and other signaling molecules for ...
    ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach
    Authors:  Junaid Akhtar, Piyush More and Steffen Albrecht, date: 02/20/2020, view: 98, Q&A: 0
    [Abstract] Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. This protocol describes a method that can be used to generate the chromatin profiles from as low as ...
    Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation
    [Abstract] Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and regenerative medicine. In this article, we describe protocols to isolate MSCs from bone marrow (BM-MSCs) and ...



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