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This detailed volume focuses on recent technological, computational, and biostatistical advances in the field of high-throughput metabolomics. Chapters encompass methods, platforms, and analytical strategies for steady state measurements and metabolic flux analysis with stable isotope-labeled tracers, in biological matrices of clinical relevance and model organisms. Mass spectrometry-based or orthogonal methods are discussed, along with computational and statistical methods to address data sparsity in high-throughput metabolomics approaches. As a part of 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, High-Throughput Metabolomics: Methods and Protocols provides tools that can bring about thenext generation of clinical biochemistry in a cost-effective, rigorous fashion, exponentially advancing our capacity to investigate nature while hastening the advent of personalized medicine.
This detailed book provides a state-of-the-art compilation of protocols relevant for the experimental study of cholestasis, each written by internationally recognized research teams that have built up long-standing expertise regarding one or more aspects of experimental cholestasis research. The first half of the collection examines methods targeted towards studying established and new mechanisms of cholestasis, while the second half explores experimental models and tools to investigate mechanisms of cholestasis and/or to predict cholestatic properties of drugs. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Experimental Cholestasis Research serves as an ideal guide for basic andapplied researchers pursuing this vital area of research in pharmacology and toxicology.
This volume explores the diversity in progenitor cell biology methods and uniquely describes techniques for isolating, generating, and characterizing progenitor cells from either tissue or embryonic stem cells. The chapters in this book cover topics such as isolating progenitor cells from sources including adipose tissue, blood, bone marrow, ear, gut, heart, pancreatic islets, and Whirton¿s jelly; deriving neural, pancreatic, and urothelial precursors from the embryonic stem cells; and culturing isolated islets in vitro to generate progenitor cells via epithelial-to-mesenchymal transition. This book also delves into digital droplet PCR; flow cytometry and cell sorting; mitochondrial assays; calcium ratiometric signaling; and 2D gel electrophoresis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, Progenitor Cells: Methods and Protocols is a valuable resource for scientists and researchers interested in learning more about this evolving and diverse field.
This volume delivers a compendium of detailed protocols to the research community in order to aid in the investigation and discovery of Yersinia virulence mechanisms using important in vivo and in vitro infection models, which have led to major advances in the field and in our understanding of Yersinia pathogen-host interactions. Beginning with a section on mouse models, the book continues with chapters covering the monitoring of bacteria during infection, invertebrate models, as well as Yersinia interaction with immune cells and immune signaling. 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, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Pathogenic Yersinia: Methods and Protocols providesa single source for researchers seeking to better understand these pathogens and the diseases they produce in humans.
This book brings together recent methods and theoretical approaches developed to dissect the activity and function of bacterial and eukaryotic Structural Maintenance of Chromosome (SMC) proteins. The protocols explore numerous subjects such as depletion systems to assess SMC function, genetic manipulation of SMC proteins, chromosomal, biochemical, and microscopy-based assays of SMC activity, as well as theoretical modeling of SMC activity. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, SMC Complexes: Methods and Protocols will greatly aid researchers looking into the mechanism of action of SMC complexes on DNA.Chapters 10 and 19 areavailable Open Access under a CC BY 4.0 license via link.springer.com.
This detailed collection gathers both established and recent technical procedures to study the Endosomal Sorting Complex Required for Transport (ESCRT) complexes in a wide range of biological systems: Archaea, A. thaliana, U. maydis, S. cerevisiae, S. pombe, C. elegans, D. melanogaster, and mammalian cells. Opening with a section on imaging techniques, the book continues with chapters covering biochemical approaches presenting strategies for production and characterization of recombinant ESCRT proteins, or of specific ESCRT protein domains, as well as genetic and proteomic experimental approaches. Written for the highly successful Methods in Molecular Biology series, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical,The ESCRT Complexes: Methods and Protocols serves as a compact guide for researchers interested in establishing an integrated approach to investigate the ESCRT machinery functions in cell biology.
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