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Neutrophil : Methods and Protocols

Neutrophils, the most abundant white cells in humans, serve as the primary cellular defense against infection. This volume provides a concise set of protocols for assessing basic neutrophil functions and investigating specialized areas in neutrophil biology. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Part I contains overviews of neutrophil biology, function, and disorders. Part II describes commonly used methods to isolate neutrophils from humans and other animal species. Part III details methods for investigating chemotaxis, transmigration, phagocytosis, and bactericidal activity. Part IV includes protocols that measure neutrophil apoptosis, calcium signal transduction, degranulation and detection of cytoplasmic granules, gene expression, transcription factors, and apoptosis. Part V provides multiple assays for measuring production of intracellular and/or extracellular reactive oxygen species, and a method that details use of the cell-free NADPH oxidase assay, an iconic assay for studies of the neutrophil NADPH oxidase. Part VI provides chapters that describe how to analyze formation and function of neutrophil extracellular traps.

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Neuronal cell death : Methods and protocols

Covers comprehensive methods on ways to assess structural and ultrastructural changes in the mitochondria, cytoskeleton, and microglia using state-of-the-art microscopy techniques including super-resolution imaging, electron microscopy, and ultra-high field MRI. The chapters in this book cover topics such as analysis of neurodegeneration in the post-mortem characterization of preclinical animal models, in vivo modeling in cell death in different model systems and brain organoids, single cell clonal analysis using Mosaic Analysis with Double Markers in genetic mouse models, and genome and proteomic methods for analysis of mRNA dynamics and quantitation of targeted peptides.

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Natural product experiments in drug discovery

Explores a wide range of evidence-based complementary medicine and various bio-analytical techniques used to define botanical products. Collecting recent work and current developments in the field of contemporary phytomedicine as well as their future possibilities in human health care, the book includes unique contributions in the form of chapters on phytomedicine and screening biological activities explained with diverse hyphenated techniques, as well as issues related to herbal medications, such as efficacy, adulteration, safety, toxicity, regulations, and drug delivery. Written for the Springer Protocols Handbooks series, chapters feature advice from experts on how to best conduct future experiments. Extensive and practical, Natural Product Experiments in Drug Discovery serves as an ideal reference for students, professors, and researchers in universities, R&D institutes, pharmaceutical and herbal enterprises, and health organizations.

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Micropropagation methods in temporary immersion systems

Presents vital techniques for the commercial micropropagation of plants. It explores numerous micropropagation protocols in different types of temporary immersion systems for plant species with agricultural, medicinal, ornamental, and forestry interest, detailing the type of bioreactor and the optimal parameters necessary to guarantee obtaining the largest number of commercial propagules. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

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Golden gate cloning : Methods and protocols

Provides an overview of the latest protocols that cover the full range of in silico, in vitro, and in vivo approaches to help researchers plan, design, and perform Golden Gate cloning experiments. The chapters in this book cover topics such as Golden Gate cloning of standardized parts; modular DNA construct design for high-throughput Golden Gate Assembly; Golden Gate cloning of Synthetic CRISPR RNA spacer sequences; Golden Gate cloning in Actinobacteria; and YeastFab cloning of toxic genes and protein expression optimization in yeast.

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Circular RNAs

Guide readers through circular RNA purification, in silico characterization, circRNA detection, sequence validation, quantification , techniques related to gain- and loss-of-function approaches, circular RNA synthesis, split ligation, engineering, nanoparticle packaging, RNA modifications on circular RNA biogenesis, RNA translation potential, and vaccines based on circular RNAs.

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Carbohydrate-protein interactions : Methods and protocols

Provides new and updated tools for studying protein-carbohydrate interactions ranging from traditional biochemical methods to state-of-the-art techniques. This book focuses on four different research themes detailing methods for screening and quantifying CAZyme activity, investigating the interactions between proteins, carbohydrate ligands, methods for the visualization of carbohydrates, protein-carbohydrate complexes, structural and “omic” approaches for studying systems of CAZymes.

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