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This second edition provides an overview of recent developments and approaches used by researchers to investigate the properties and functions of mammary epithelial and stem cells, which will contribute to understand the heterogeneity of the mammary gland and of breast cancer. Chapters detail processes used to characterize stem cells, single cell RNA sequencing, computational methods, sophisticated imaging techniques, and a variety of model systems, among others. 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. Authoritative and cutting-edge, Mammary Stem Cells: Methods and Protocols, Second Edition aims to make available protocols used to navigate the intricate behavior of mammary stem cells and to gain further knowledge to take us closer to the design of innovative strategies to prevent and treat breast cancer.

This volume presents an all-inclusive overview of molecular biological methods to study ABA signaling transport, and metabolism in several plant species. The chapters in this book cover topics such as biochemical and molecular biological characterization of ABA receptors; assay systems for ABA transporters; quantification and visualization of ABA levels; and comprehensive omics 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. Cutting-edge and thorough, Abscisic Acid: Methods and Protocols is a valuable resource for scientists and researchers who are interested in learning more about this developing field.

This detailed volume explores techniques for researching brown adipose tissue (BAT) and the fascinating biology and therapeutic potential of thermogenic adipocytes. The content reflects the advancing technologies in genetics, imaging, and 'omics strategies that are allowing researchers to probe BAT biology at unprecedented depths and detail, yet it also presents classic physiology principles, which remain the core tenets of BAT biology. 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, Brown Adipose Tissue: Methods and Protocols provides perspectives and detailed protocols for the benefit of both new BAT researchers looking for guidance as well as seasoned researchers who would like to expand their toolkits. Chapter 12 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume explores the latest techniques used to study brain function and pathophysiology of major depressive disorder (MDD), and includes suggestions of new therapeutic approaches for the treatment of MDD. The chapters into this book are organized into five parts. Part One discusses advanced approaches to studying well-established pathophysiological mechanisms. Part Two details behavioral research methods for MDD. Part Three looks at the cellular and molecular research methods for major depression, and Part Four describes the latest developments in non-invasive neuroimaging. Part Five focuses on the pharmacological and non-pharmacological interactions, including antidepressant agents and their properties, such as sexual side effects and neuroimaging biomarkers. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory.Cutting-edge and comprehensive, Translational Research Methods for Major Depressive Disorders is a valuable resource for researchers and scientists interested in learning more about this important and developing field.

This volume provides computational methods and reviews various aspects of computational studies of protein aggregation. Chapters discuss the relationship between protein misfolding and protein aggregation, methods of prediction of aggregation propensities of protein, peptides, protein structure, results of computer simulations of aggregation, and computational simulations focused on specific diseases such as Alzheimer's, Parkinson's, and preeclampsia. 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. Authoritative and cutting-edge, Computer Simulations of Aggregation of Proteins and Peptides aims to ensure successful results in the further study of this vital field.

This volume explores detailed methods to investigate various aspects of biology related to cell polarity, or asymmetry within a cell. Molecular, cellular, and tissue-level regulation and function as well as diseases caused by impairment of cell polarity are explored by these methods. Beginning with advanced imaging and biochemical methods, the book continues with planar cell polarity (PCP) signaling in morphogenesis in diverse developmental contexts, apicobasal (AB) cell polarity in development and diseases, as well as directional cell migration and biomechanics in cell polarity. The collection includes the usage of a wide variety of model systems and an extensive array of techniques, including genetic, imaging, biochemical, and biomechanical. Written for the highly successful Methods in Molecular Biology 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 comprehensive, Cell Polarity Signaling: Methods and Protocols aims to enable researchers to delve into the stimulating field of cell polarity and contribute to our understanding of how coordinated control of protein stability, trafficking, membrane retention, post-translational modification, and dynamic organization leads to active regulation of cell polarity. Chapter 29 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This detailed volume explores hepatic stellate cells, which have become a spotlight of liver cell biology due to their pleiotropic functions that extend well beyond extracellular matrix production to pivotal roles in hepatic homeostasis, immunity, and metabolism. The book features methods to isolate, evaluate, and manipulate this cell type in an effort to elucidate hepatic biology and establish prospects for treating disease. 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, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Hepatic Stellate Cells: Methods and Protocols serves as an ideal aid to help researchers accelerate the outstanding science that has steadily unveiled the mysteries of stellate cell biology and their role in disease.

This volume provides both experienced and new microscopists with methods and protocols to perform fluorescence microscopy-based experiments. The book is divided into four parts detailing basic fluorescent microscopy, quantitative methods, imaging living animals, human tissue samples, approaches for imaging at a near-molecular level, and approaches to image analysis. 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, Fluorescent Microscopy aims to be a useful practical guide to researches to help further their study in this field.

This volume explores a collection of different protocols for the analysis and characterization of DNAzymes and their functions. The topics covered in this book range from bioinformatics and molecular dynamics simulations for the study or modification of nucleic acids to the descriptions of spectroscopic, fluorescence-based, or crystallographic methods to understand the structure and function of DNAzymes. 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, DNAzymes: Methods and Protocols is a valuable resource for scientists and researchers interested in learning more about this evolving field.

This volume provides detailed, up-to-date methods used in research on Atherosclerosis. Chapters guide readers through an overview of the pathogenesis of atherosclerosis and model systems together with in vitro, ex vivo, in vivo and emerging methods in atherosclerosis research. 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. Authoritative and cutting-edge, Atherosclerosis: Methods and Protocols serves as an invaluable resource for those engaging in research on atherosclerosis and cardiovascular disease, as well as for researchers who are new to the field and students at both the undergraduate and postgraduate level.

This volume contains a comprehensive collection of laboratory protocols used by researchers to analyze varied aspects of non-alcoholic steatohepatitis (NASH). The chapters in this book cover topics such as methods for histological diagnosis of NASH; the purpose of generating an in vivo NASH model; protocols for isolating hepatocytes and Kupffer cells, bone marrow derived macrophages, and adipocytes; techniques to develop human pluripotent stem cells-derived liver organoids; single-cell and RNA-sequencing; and a description of how to extract exosomes and exosomal miRNAs from mesenchymal stem cells. 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. Thorough and practical, Non-Alcoholic Steatohepatitis: Methodsand Protocols is a valuable resource for new and experiences investigators studying NASH, and serves as an essential reference on NASH for basic and clinical researchers and students.

This fully updated book explores all-new and revised protocols involving the use of in silico models, particularly with regard to pharmaceuticals. Divided into five sections, the volume covers the modeling of pharmaceuticals in the body, toxicity data for modeling purposes, in silico models for multiple endpoints, a number of platforms for evaluating pharmaceuticals, as well as an exploration of challenges, both scientific and sociological. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detail and implementation advice necessary for successful results. Authoritative and comprehensive, In Silico Methods for Predicting Drug Toxicity, Second Edition aims to guide the reader through the correct procedures needed to harness in silico models, a field which now touches a wide variety of research specialties.

The latest developments in the area of microbial immunity are explored and explained in this timely volume, including new conceptual frameworks for the processing and presentation of peptides and nonpeptide antigens from microbes to T cells. Such experimental models have been instrumental in defining some of the pathways for generating protective immune responses against microbes. Other topics covered include: DAP 12, CD4+, Th1 and Th2 Cells, HIV infection, fusion, and tropism, and vaccine development.

This second edition includes a collection of antigen production and delivery strategies for vaccine development in veterinary species. New and updated chapters guide readers through protocols for antigen production, experimental antigen delivery and the analysis of immune responses upon vaccination. 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, Vaccine Technologies for Veterinary Viral Diseases: Methods and Protocols, Second Edition aims to be a useful and practical guide to researches to help further their study in this field.

This fully updated collection includes chapters on a wide array of techniques that are vital for advancing our understanding of the physiological and pathological effects of estrogen, mediated by estrogen receptor a (ERa) and estrogen receptor ß (ERß) as well as the non-genomic membrane-bound G-protein coupled estrogen receptor (GPER/GPR30). The protocols range from standard methods and important laboratory workhorses such as immunoblot and RT-qPCR, to newer technologies such as PET imaging, reporter animal models, polysome fractionation, and many others, involving a broad range of tissue types to demonstrate the variety of estrogen receptor effects. Written for this 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, Estrogen Receptors: Methods and Protocols, Second Edition serves as an ideal guide to researchers working to explore the vibrancy and excitement of the field of estrogen receptors and estrogen actions.

This book provides protocols on the preparation of synaptic vesicles from different model systems and provides instructions for functional investigations of their transport and formation, scaffolding, filling and priming, release, recycling, and degradation. Chapters guide readers through computational approaches, simulating different aspects of synaptic vesicle anatomy and function. 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, Synaptic Vesicles: Methods and Protocols aims to be a useful practical guide to researchers to help further their study in this field.

This volume provides detailed technical protocols on current biomedical technologies and examples of their applications and capabilities. Chapters focus on molecular and cellular analytical methods, experimental new drug delivery approaches, guided surgery, implants and tissue engineering. 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, tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and practical, Biomedical Engineering Technologies, Volume 2 provides technical details in descriptions of major technologies by experts in the field.

This second edition volume expands on the previous edition with updates about the latest state-of-the-art techniques used in leading hemostasis and thrombosis laboratories for diagnosis and exclusion of hemorrhagic and thrombotic diseases. The chapters in this book are organized into seven parts. Part One provides a general overview on hemostasis and thrombosis, preanalytical issues in testing, and routine hemostasis assays. Part Two covers laboratory testing for thrombophilia, including reviews for activated protein C resistance, protein C, lupus anticoagulant testing, and antiphospholipid antibodies. Part Three addresses monitoring continuous anticoagulant infusions and measuring the effects of oral anti-thrombotic therapy. Part Four talks about heparin induced thrombocytopenia and vaccine induced immune thrombotic thrombocytopenia. Part Five and Six cover ADAMTS13 activity testing and new information on bleeding disorders such as chromogenic factor VIII assays, measurement of emicizumab, and treatment of hemophilia A and B. Finally, Part Seven discusses global assays, research applications, and postanalytical considerations. 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, Hemostasis and Thrombosis: Methods and Protocols, Second Edition is a valuable resource for scientists and researchers struggling to identify the appropriate methods for hemostasis and thrombosis testing, or who seek additional expert guidance on such testing.

This third edition provides updated and new chapters on recent innovations in basic renal research. Chapters are divided into three sections covering in vitro, in vivo, and ex vivo models of kidney disease, recent advances in imaging techniques and protocols for performing analytical and functional measurements in the kidney; techniques that are both topical and of widespread relevance to the study of kidney biology and disease. 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 methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Kidney Research: Experimental Protocols, Third Edition aims to be comprehensive guide for researchers in the field.

This volume presents a broad collection of state-of-the-art methods to study the biology of thermogenic fat using in vitro cell culture and animal models. Chapters guide the readers through protocols on differentiation of human pluripotent stem cells and murine adipocyte precursors; methods for measuring mitochondrial respiration, heat generation, brown fat activation, and effects on energy metabolism in mice; and techniques for AAV-mediated gene delivery, transplantation of adipose tissue, isolation of adipose tissue immune cells and extracellular vesicles, and mass spectrometry-based profiling of brown fat lipids. 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 methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge,Thermogenic Fat: Methods and Protocols aims to be comprehensive guide for researchers in the field.

This volume covers current and emerging techniques for studying single-domain antibodies (sdAbs). Chapters guide readers through the biology and immunology of sdAbs in camelids and sharks, isolation of sdAbs, protein engineering approaches to optimize the solubility, stability, valency and antigen binding affinity of sdAbs, and specialized applications of sdAbs. 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, Single-Domain Antibodies: Methods and Protocols aims to be a useful, practical guide to help researchers further their studies in this field.

This volume provides an up-to-date collection of protocols describing some of the key methods to investigate the integrated stress response (ISR), a vital evolutionarily conserved mechanism that enables eukaryotic cells to adapt to stress conditions and alter their gene expression programs. The content of the book is split between techniques to analyze mRNA translation regulation and methods to analyze interaction networks and ribonucleoprotein (RNP) granules. 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, The Integrated Stress Response: Methods and Protocols serves as an ideal guide to help accelerate research into the complex and fascinating biology of the ISR.

This detailed volume aims to elucidate the molecular mechanisms that underlie antifungal resistance. The book highlights methods to identify and characterize antifungal activity, to define and characterize strains with altered responses to antifungal drugs, to investigate the genetic and molecular mechanisms of these alterations of antifungal drug susceptibility, and, finally, to approach the study of these processes in animal models of fungal infection. 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, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Antifungal Drug Resistance: Methods and Protocols serves as an ideal resource for the field and a guide for investigators to study this important translational aspect of fungal biology.

This detailed collection focuses on current methodologies for studying various crop diseases. The book explores phytopathogen detection, host-pathogen interactions at the earliest stages of colonization, RNA-seq analyses, as well as host responses downstream of, or coincidental to, gene expression and a newly established protocol for gene editing of Medicago sativa. 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, Plant-Pathogen Interactions serves as an ideal guide for researchers working in the vital area of plant diseases.

This volume explores the latest integrated bioprocesses and technologies used to study the production of the target recombinant protein of therapeutic or diagnostic interest; its isolation, purification, and stabilization; and the bio-interaction and structural analyses. The chapters in this book are organized into four parts. Part One covers production methods of soluble and membrane proteins in prokaryotic and eukaryotic expression systems, such as Lactococcus lactis and Escherichia coli. Part Two describes traditional and novel approaches for recombinant protein purification and stabilization, and buffers and additives. Part Three discusses automated methods in structural biology based on in silico approaches; and Part Four provides examples of advanced protein investigation methodologies to assess structural analysis such as high-throughput protein crystallization and time-resolved serial crystallography. 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, Advanced Methods in Structural Biology is a valuable resource to those in academia (i.e., graduate students and postdoctoral researchers) and researchers in the pharmaceutical industry who wish to learn more about this developing field. Chapter 5 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This detailed new edition compiles fully updated practical methodologies and approaches for diagnosticians and researchers working toward a better understanding of animal coronaviruses. The book explores vital techniques in virus isolation, diagnostic approaches via the detection of viral nucleic acids or proteins, serology, disease-targeted animal species models, and next-generation sequencing and bioinformatics. Written for the invaluable Springer Protocols Handbooks series, chapters contain readily reproducible laboratory protocols as well as expert tips on troubleshooting and avoiding known pitfalls. Thorough and hands-on, Animal Coronaviruses, Second Edition will help readers design and carry out their increasingly important projects in clinical research, diagnostics, disease surveillance, and epidemiological study involving this family of viruses.