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This volume provides detailed protocols for the identification and understanding of vasculogenic mimicry process in vitro and in vivo, in addition to protocols for microscopy and histology. Chapters guide readers through different materials, commercial and homemade scaffolds, Matrigel, cancer spheroids, 3D tissue constructs, vasculogenic processes, and mathematical model building. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Vasculogenic Mimicry: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. 

This fully updated volume explores improved and extended protocols for embryonic stem cell (ESC) research, provided with the most direct and informative methodologies currently available. The book examines how these models for cell lineage and differentiation studies have continued to mature into a critical research workhorse. Written for 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Embryonic Stem Cell Protocols, Fourth Edition serves as an ideal resource for researching mining the depths of the ESC field.

This volume provides an overview of well-established methods optimized for diverse archaeal model organisms and is a source of protocols facilitating access to the molecular and cellular biology characterization of these fascinating organisms. Chapters are divided into five parts detailing available genetic tools, molecular and cellular biology methods, strategies to study the ecophysiology of archaea, and classroom protocol. Each main thematic part is also introduced by future-oriented and authoritative primers. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.Authoritative and cutting-edge, Archaea: Methods and Protocols aims to be a foundation for future studies and to be a source of inspiration for new investigations in the field.

This volume presents a collection of protocols to study effector-triggered immunity (ETI) in both plants and animals from eminent groups in the field. The chapters in this book cover topics such as genetic manipulation of plant and animal pathogens, host cells, and the analysis of key host responses; and techniques used for the analysis of inflammasome activation, cell death pathways, and mitochondria damage in response to pathogens. All of these topics cover a broad spectrum of immunological, biochemical, cell biological, and structural biology approaches to examine ETI. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and practical, Effector-Triggered Immunity: Methods and Protocols is a valuable resource for both expert and novice researchers who are interested in learning more about the important and developing field of ETI.

This volume provides an overview of methods for plant species, mainly species with high economic value, to achieve in vitro propagation through somatic embryogenesis. Chapters guide readers through somatic embryogenesis protocols, somatic embryos in plants, and new methodologies that are used out to solve in vitro recalcitrance of many species to this morphogenetic route. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Somatic Embryogenesis: Methods and Protocols aims to guide future researcher to achieve success in obtaining and regenerating somatic embryos. Chapter 6 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume provides updated protocols for chemical protein synthesis. Chapters guide readers through development methods, strategies, and applications of protein chemical synthesis. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Chemical Protein Synthesis aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. 

This detailed volume explores a variety of cutting-edge techniques used to interrogate spatial genome organization. Beginning with a section covering the vital chromosome conformation capture (3C) technique, this collection continues with chapters on targeted Hi-C approaches, sequencing-based approaches to assess nuclear environment, as well as single-cell technologies to better characterize the heterogeneity and dynamics of nuclear architectures and approaches to visualize them by microscopy. Finally, in order to be able to ask functional questions about the role of spatial chromatin organization in genomic control, the last section provides methods for acute manipulations of chromatin architecture. 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, Spatial Genome Organization: Methods and Protocols is an ideal resource for researchers searching for the best techniques to address their own specific research questions.

This volume discusses the latest techniques used to identify cancer drug resistance determinants at the molecular, cellular, and functional levels. Chapters in this book cover up-to-date topics including tumor-microenvironment cell co-culture methods and microfluidics systems; workflows for functional assessment of drug resistance in vitro and in vivo; quantitative techniques for identifying quiescent blood-flow circulating cells; and single-cell characterization methods, such as mass cytometry. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and practical, Cancer Drug Resistance: Methods and Protocols is a valuable resource for all scientists and researchers who are looking to learn more about the latest developments in understanding and overcoming anticancer drug resistance.  

This volume discusses protocols that cover synthesis, screening by selection, and analysis of DNA-encoded chemical libraries (DEL). Chapters in this book focus on methods used to practice DEL technology and include solution phase library synthesis using a variety of chemistries; DNA encoding of chemical structure; design, preparation and analysis of target proteins and tool compounds; screening of soluble protein targets by affinity selection; DEL qPCR, preparative PCR and DNA sequence analysis; computational methods used to analyze selections and choose compounds for resynthesis; and analysis of hit compounds. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, DNA-Encoded Chemical Libraries: Methods and Protocols is a valuable resource for scientists interested in DEL technology for drug discovery, and will contribute to the continued advancement in this important field.

This updated volume explores Candida and candidiasis methods, useful to a wide variety of Candida scientists including those new to the field. Beginning with a section on the Candida genome, the book continues by covering techniques for antifungal discovery and studying resistance, pathogenesis and virulence, communities, as well as immunity. Biofilm protocols are also featured. 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 up-to-date, Candida Species: Methods and Protocols, Second Edition serves as an ideal guide for researchers working to further our understanding of this family of infective fungi.

This volume discusses methods used in the fields of molecular and cellular biology for detecting and studying cell death, especially in cancer and cancer therapy. Chapters in this book cover topics such as non-destructive, real-time Annexin V apoptosis assay; assessment of the immune response to tumor cell apoptosis and efferocytosis; mass cytometry assessment of cell phenotypes and signaling states in human whole blood; cell cycle analysis of ER stress and autophagy; and simultaneous detection of inflammasome activation and membrane damage during pyroptosis. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and practical, Apoptosis and Cancer: Methods and Protocols is a valuable resource and practical guide for both novice and expert researchers looking into the "e;meaning of death."e;

This volume details experimental protocols to study structure and function of hepatocytes in vivo and in vitro, and for characterizing heterogeneity and plasticity of hepatocytes in healthy and in injured liver. Chapters guide readers through separating hepatocyte subfractions, characterizing the heterogeneity, novel culture systems, hepatocyte-like cells from human pluripotent stem cells, analyzing drug metabolism, monitoring protein sorting, manipulating gene expression, and methods for expanding primary hepatoblastoma and identifying hepatocellular carcinoma subtypes. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Hepatocytes: Methods and Protocols aims to provide techniques to further understand molecular mechanisms involved in the regulating hepatocyte functions and underlying liver diseases.

This updated volume provides stepwise instructions for the analysis of numerous clinically important analytes by mass spectrometry. Mass spectrometry offers clinical laboratory scientists a number of advantages including increased sensitivity and specificity, multiple component analysis, and limited need for specialized reagents. These techniques are essential in laboratory fields including endocrinology, biochemical genetics, drug analysis, proteomics, and pathogen identification. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Clinical Applications of Mass Spectrometry in Biomolecular Analysis: Methods and Protocols, Second Edition is an ideal resource for clinical laboratory scientists who are already using or thinking of bringing mass spectrometry to their laboratories.

This new edition offers a state-of-the-art and integrative vision of pharmacogenomics by exploring new concepts and practical methodologies focusing on disease treatments, from cancers to cardiovascular and neurodegenerative disorders and more. The collection of these theoretical and experimental approaches facilitates problem-solving by tackling the complexity of personalized drug discovery and development. Written by leading experts in their fields for the highly successful Methods in Molecular Biology series, the book aims to provide across-the-board resources to support the translation of pharmacogenomics into better individualized health care. Authoritative and up-to-date, Pharmacogenomics in Drug Discovery and Development, Third Edition aims to aid researchers in approaching the challenges in pharmacogenomics and personalized medicine with the introduction of these novel ideas and cutting-edge methodologies.

This volume provides the most widely used protocols and techniques employed in lipopolysaccharide( LPS) transport studies. Chapters detail identification and analysis of essential genes and crucial mutations in LPS transport-related genes, LPS profile determination, LPS chemical analysis, protein-LPS interaction, and LPS visualization. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.Authoritative and cutting-edge, Lipopolysaccharide Transport: Methods and Protocols aims to deliver an overview on the multidisciplinary study of LPS transport and to identify new and innovative antibacterial strategies.

This detailed volume presents a series of protocols that are representative of recent developments and improvements in induced pluripotent stem cells (iPS cells) and corresponding human disease models. Reflecting the latest technology for generating induced pluripotent stem cells (iPS cells) and their initial characterization, the book explores techniques invaluable both for studies of disease-specific cell types and for their potential applications in regenerative medicine. 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 and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Induced Pluripotent Stem Cells and Human Disease: Methods and Protocols serves as a vital guide that is valuable for not only experts but also novices in the stem cell field.

This volume covers the latest techniques that study the synthesis of melatonin, its receptor function, and its effects at the cellular and systemic level. The chapters are organized into three parts. Part One describes methods for the detection of melatonin and its biological derivatives in various biological samples, the manipulation of melatonin synthesis by the pineal gland in animals, and the principal source of melatonin in mammals. Part Two explores methods to measure the biological effects and consequences of melatonin binding to high-affinity G protein-coupled receptors. Part Three describes methods to measure the physiological effects that are regulated by melatonin in animals, particularly in rodent models. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, Melatonin: Methods and Protocols is a valuable resource for any researcher interested in investigating melatonin, from its production to its mechanisms of action and systemic effects.  

This volume details state-of-the- art methods on computer-aided antibody design. Chapters guide readers through information on antibody sequences and structures,  modeling antibody structures and dynamics, prediction and optimization of biological and biophysical properties of antibodies, prediction of antibody-antigen interactions, and computer-aided antibody affinity maturation and beyond. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.Authoritative and cutting-edge, Computer-Aided Antibody Design aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. Chapter 2 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume provides protocols for computational, statistical, and machine learning methods that are mainly applied to the study of metabolic engineering, synthetic biology, and disease applications. These techniques support the latest progress in cross-disciplinary research that integrates the different scales of biological complexity. The topics covered in this book are geared toward researchers with a background in engineering, computational analytical, and modeling experience and cover a broad range of topics in computational and machine learning approaches. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Comprehensive and practical, Computational Biology and Machine Learning for Metabolic Engineering and Synthetic Biology is a valuable resource for any researcher or scientist who wants to learn more about the latest computational methods and how they are applied toward the understanding and prediction of complex biology. 

This thorough volume explores protocols of proteome- and metabolome-wide strategies for the identification of protein-small molecule complexes in different organisms, in order to shed light on these important regulatory interactions. Experimental and computational strategies to characterize protein-metabolite interactions are discussed, and recent advances in enabling technologies are featured as well. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detail and expert implementation advice to ensure success in future research. Authoritative and practical, Cell-Wide Identification of Metabolite-Protein Interactions will aid researchers seeking a better understanding of the mechanisms of signal transduction occurring in the cell and assessing the effect of complex formation on cell physiology.

This volume details applications in molecular biological techniques and focuses on applications to determine the involvement of glycans in virus interactions. Chapters guide readers through glycan analysis, glycan distribution analysis, glycan and lectin microarray, preparation of recombinant viral domain protein, reverse genetics and receptor binding, virus-host interactions and receptor binding, and sialidase: assays and inhibitors. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Glycovirology: Methods and Protocols aims to be an essential resource for researchers who wish to learn more about glycovirology. 

This detailed book examines the main methods to study mammalian monoamine oxidases (MAOs), ranging from cell biology to computational chemistry. Beginning with techniques on how to obtain pure samples of MAO A and MAO B, the volume continues by covering assays and techniques used to measure MAO enzymatic activity and perform inhibition studies, methods to address cellular localization and function of MAOs, either in cell lines or in animal models, as well as computational methods applied to rational drug design approaches that are used to develop new MAO inhibitors. Written for 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Monoamine Oxidase: Methods and Protocols serves as a vital resource for scientists who are interested in studying MAOs and other similar amine oxidase enzymes.

This volume provides methods and models for investigating the immunosuppressive subset of CD4+ T-cells, regulatory T-cells (Treg). Chapters guide readers through, basic protocols for analyzing Treg in mice and humans, single-cell analysis for analyzing Treg, and methods for the analysis of Treg in clinical application. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Regulatory T-Cells: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. 

 This volume details the history of Retinitis Pigmentosa and current treatment options. Chapters guide readers through CRISPR, gene therapy, stem cell therapy, next-generation sequencing methods, gene editing, and translational applications of other therapies to the treatment of Retinitis Pigmentosa. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Retinitis Pigmentosa aims to be a useful practical guide to researches to help further their study in this field.  

This volume provides a comprehensive reference for researchers aiming to bring new techniques and approaches to their scientific research using urodeles. Chapters are authored by leaders in the field and meant to guide readers through laboratory colony husbandry, traditional molecular techniques, experimental manipulation and surgeries, bioinformatics and genomics, transgenics and lineage-tracing, and physiological and organismal techniques. In addition to laboratory methods, this volume highlights techniques developed for field studies and work with wild-caught animals. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and state-of-the-art, Methods in Salamander Research aims to be a practical guide for researchers interested in deploying new  methodologies in their lab or in the field. 

This volume provides readers with a broad collection of theoretical, computational, and experimental methods to quantitatively study the properties of phase-separate biomolecular condensates in diverse systems. The chapters in this book cover topics such as theoretical and computational methods; methods for in vitro characterization of biomolecular condensates; and techniques that enable in-cell characterization of biomolecular condensates. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and expert tips on troubleshooting and avoiding known pitfalls.Comprehensive and thorough, Phase-Separated Biomolecular Condensates: Methods and Protocols is a valuable resource that helps researchers learn and use established methods to study both biophysical properties and biological functions of biomolecular condensates. 

This volume brings together cutting-edge laboratory protocols to characterize the novel fluorescent proteins (FPs) and approaches based on fluorescent proteins that aim to answer some of the key cell biological questions. The book covers topics ranging from the database of fluorescent proteins to their characterization and adaptation to a wide range of biological systems. 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. Authoritative and practical, Fluorescent Proteins: Methods and Protocols serves as an ideal guide for students and academicians enthusiastic about the recent progress in the practical application of fluorescent protein technology.

This volume covers the most up-to-date methods and techniques used to further the understanding of chromaffin cell biology and pharmacology. Chapters guide readers through the basic mechanisms that regulate the stimulus-secretion coupling, chromaffin, tumor-derived cell PC-12 , morphology, biochemistry, pharmacology, electrophysiology, and electrochemistry. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Chromaffin Cells: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field. 

This volume details histochemical techniques for the detection of specific molecules or metabolic processes, both at light and electron microscopy. Chapters are divided into seven sections covering Vital histochemistry, Carbohydrate histochemistry, Protein histochemistry, Lipid histochemistry, Nuclear histochemistry, Plant histochemistry and Histochemistry for Nanoscience. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. The volume also contains three discursive chapters on Histochemistry in advanced cytometry, Lectins and Detection of molecules in plant cell walls by fluorescence microscopy. Authoritative and cutting-edge, Histochemistry of Single Molecules: Methods and Protocols, Second Edition aims to be a useful practical guide for researchers to help further their study in this field.

This detailed volume collects updates on the technical advances in hematopoietic stem cell research and incorporates new techniques focused on the molecular/genetic, cellular, and whole organism levels. Exploring methods that apply stress to hematopoiesis, the book also contains chapters focused on better understanding the role of hematopoietic niches and their cellular components, as well as in vivo models that test and quantitate stem cell function and are key to further development of therapeutic applications. 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. Authoritative and up-to-date, Hematopoietic Stem Cells: Methods and Protocols serves as a valued addition to laboratories focused on understanding hematopoietic stem cell biology and the therapeutic advances that can be derived from it.