Books in the Methods in Molecular Biology series

This volume covers protocols related to both pluripotent and somatic stem cells, including the ethical procurement of tissues and cells for the provision of ¿seed stock,¿ standardized methods for deriving hESCs and iPSCs, isolating mesenchymal stem cells, cell culture and cryopreservation, in addition to quality assurance and information management. Stem Cell Banking: Concepts and Protocols aims to contribute to the development of this field by providing information that is essential to establishing a bona fide stem cell bank. 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, Stem Cell Banking: Concepts and Protocols is a valuable resource for stem cell scientists and novices to the field, and will help strengthen and maximize their use of existing and future stem cell resources.

This second edition provides updated and new chapters on T-Cell trafficking. In addition to detailed experimental procedures, the interested reader will find informative introductory chapters on the relevance of T-Cell trafficking in thymic population and maturation, traffic through secondary lymphoid organs during ¿physiological¿ resolving inflammation and during immune responses, as well as T-Cell trafficking in chronic inflammatory diseases. Importantly, chapters cover methods from in silico modeling of cellular interactions, in vitro adhesion assays, through ex vivo functional assays to integrated intravital modeling of T-Cell trafficking through organs. Written in the highly successful Methods in Molecular Biology series format, each methods chapter includes a short introduction to the topic, 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, T-Cell Trafficking: Methods and Protocols, Second Edition aims to be an essential point of reference for those new to the field of T-Cell trafficking, or to those looking to expand their technical capabilities.

This volume provides laboratory protocols essential for studies on lysosomal biology. Chapters aim to guide researchers in their exploration of lysosomes, both under normal conditions and in pathological processes. 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 practical, Lysosome: Methods and Protocols aims to provided protocols that will guide and inspire further research and generate new insights into this fascinating organelle.

This volume provides easily accessible and comprehensive collection of methods, techniques, and strategies to investigate the molecular and cellular biology of peroxisomes in different organisms. Chapters detail valuable instructions, guidelines and protocols for molecular cell biologists, biochemists and biomedical researchers with an interest in peroxisome biology. Chapters in Peroxisomes: Methods and Protocols illustrate the isolation of peroxisomes, investigation of properties of membrane proteins, protocols to investigate and manipulate peroxisomes in cellular systems, detection of peroxisomes, including immunofluorescence, cytochemistry, cryo-immuno electron microscopy, and live cell imaging approaches.Authoritative and practical, Peroxisomes: Methods and Protocols aims to be useful for those already working on peroxisomes as well as for those who would like to start working on this fascinating organelle.

Beginning with general strategies, the volume continues with coverage of peptide switches, fluorescent and bioluminescent sensors, -lactamase sensors, proteolytic sensors, optogenetic switches, as well as cellular signaling switches.

This volume provides readers with a better understanding of organogenesis in developmental biology and next-generation organ regenerative therapy. This book focuses on recent studies of organ regeneration from stem cells using in vitro 3D cell culture and manipulation. The chapters cover topics such as generation of a 3D retinal tissue formation; functional salivary gland regeneration; liver regeneration using cultures liver bud; and in vivo model of small intestine. 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 thorough, Organ Regeneration: 3D Stem Cell Culture and Manipulation is a valuable resource for scientists and researchers who are interested in this field.

Ataxiätelangiectasia (A-T) is a rare and severe genetic disorder affecting children. A-T is a multisystem disease characterized by progressive neurodegeneration, immunodeficiency and cancer predisposition. This detailed volume explores the ever expanding field of research into the ATM (ataxia-telangiectasia, mutated) gene and the role played by ATM kinase in DNA damage signaling and diverse cellular processes. What follows is a handy desktop reference for both seasoned A-T researchers and postgraduate students, as it demonstrates the breadth of recent developments in A-T studies. 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. Both classic and cutting-edge techniques are described, including ATM gene mutation detection, assays for radiosensitivity and radioresistant DNA synthesis, new methods to measure ATM kinase activity by imaging microscopy and high content screening as well as proteomics, phosphoproteomics and bioinformatics approaches to decipher ATM-dependent signalling pathways. Additional methods include generation of patient-specific stem cells and approaches to study ATM functions in the nervous system. Comprehensive and practical, ATM Kinase: Methods and Protocols aims to ignite and attract the interest of colleagues from diverse fields to A-T research in an effort to bring their expertise and fresh ideas to resolve many A-T puzzles still waiting to be pieced together and to alleviate the suffering of A-T children and their families.

This second edition volume expands on the previous editions with detailed protocols on (cyano) bacterial and mold fungus toxins, and a specific focus on Gram-negative endotoxins (lipopolysaccharides).

This volume provides an overview of zygotic genome activation (ZGA) and the use of recent tools to elucidate the events that occur during ZGA.

This second edition provides an update on technology and an accelerated tutorial to assist, students, entrepreneurs, new investigators, and established investigators who want to quickly become versed in, and immersed in, the entire process from discovery to clinical trial validation and commercial public benefit. Chapters aim to cover the full spectrum of molecular profiling from tumor staging and grading through biomarker discovery, to commercialization. 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 practical, Molecular Profiling: Methods and Protocols, Second Edition aims to be a useful guide on molecular profiling.

This book presents multiple new and classical methods for studying the vital poly-ADP-ribose (pADPr) pathway. Beginning with techniques for the detection and quantification of the product of poly(ADP-ribose) polymerase (PARP) enzymatic activity and detection of variation in pADPr production during the cell cycle, the volume continues with sections on the identification of pADPr protein acceptors, methods focusing on studying molecular mechanisms of PARP functions in eukaryotic cells, particularly those involved in control of DNA repair and oxidative stress, as well as in expression regulation, approaches to the in vitro reconstitution of PARP-1 interaction with chromatin, the development and testing of small molecule PARP inhibitors, and the functions of understudied members of PARP family. 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, Poly(ADP-Ribose) Polymerase: Methods and Protocols, Second Edition serves as an ideal companion to the first edition for scientists whose investigations involve this important pathway.The chapter 'Identifying and Validating Tankyrase Binders and Substrates: A Candidate Approach' is published open access under a CC BY 4.0 license.

This volume provides methods for modern macromolecular crystallography, including all steps leading to crystal structure determination and analysis. 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 practical, Protein Crystallography aims to ensure successful results in the further study of this vital field.

This volume provides protocols that revolve around three pillars of progress in the plant genomics field: genotypes, phenotypes, and the molecular processes in between. Chapters in Plant Genomics: Methods and Protocols are not restricted to the predominant model species Arabidopsis thaliana, hoping to encourage and facilitate other researchers to expand their research to other species. 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 practical, Plant Genomics: Methods and Protocols aims to serve as an inspiration for further studies in plant genomics.

This book provides an overview of established 3D cell culture assays from leaders in the field. Their contributions cover a wide spectrum of techniques and approaches for 3D cell culture, from organoid cultures through organotypic models to microfluidic approaches and emerging 3D bioprinting techniques, which are used in developmental, stem cell, cancer, and pharmacological studies, among many others. 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. Comprehensive and cutting-edge, 3D Cell Culture: Methods and Protocols aims to inspire researchers to develop novel 3D cell culture techniques according to their specific scientific needs and interests, leading to a new generation of physiologically relevant and realistic 3D cell cultures.Chapter 15 of this book is available open access under a CC BY 4.0 license.

This detailed volume focuses on the development of mycosis vaccines via the most common etiological agents of mycoses, such as Aspergillus and Candida species, followed by Cryptococcus, dimorphic fungi, and members of Mucoraceae family of fungi.

This detailed collection provides an accessible compendium of up-to-date methods focused on the study of RNAi and small regulatory miRNAs in stem cells. Beginning with a brief introductory section, the volume continues by exploring methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, recently developed methods for miRNA expression and functional analysis, as well as usage of CRISPR/Cas 9 to knockout an individual gene for functional studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, list of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, RNAi and Small Regulatory RNAs in Stem Cells: Methods and Protocols aims to accelerate progress in this crucial field by reducing the time required to decipher and put into practice procedures published in the literature.

This detailed volume examines the latest techniques and protocols associated with zymography, the practice of detecting enzyme activity on electrophoretic gels. Sections cover subjects such as endopeptidase zymography, reverse zymography & in situ zymography, 2D zymography, as well as a variety of special cases.

This volume presents a collection of computational and experimental protocols pertaining to the creation, characterization, and utilization of RNA nanostructures.

In this present volume, different approaches are detailed to produce membrane proteins, purify them, study their function, determine their structure, and model them in membrane.

This two-volume book is a valuable resource to students, researchers, scientists, commercial producers, consultants and policymakers interested in agriculture or plant sciences particularly in date palm biotechnology. Chapters in Date Palm Biotechnology Protocols: Volume 2: Germplasm Conservation and Molecular Breeding guides readers through methods and protocols on germplasm in vitro conservation, molecular analysis of in vitro cultures, genetic diversity, cultivar identity, gender identification, genomics, and proteomics. 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 practical, Date Palm Biotechnology Protocols: Volume 2: Germplasm Conservation and Molecular Breeding aims to supplement the previous volume and to provide precise stepwise protocols in the field of date palm biotechnology.

This volume presents a comprehensive collection of quick assays for the detection of nuclear and mitochondrial DNA damage and its effects in live and fixed cells and tissues, and in bacterial genomes.

This book expands on the previous volumes with new chapters focusing on functional characterization of phage and their proteins, and on the development of phage therapy by outlining novel models.

This volume covers methods for determination of autoantibodies in rheumatic connective tissue diseases and organ-specific diseases. 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, Autoantibodies: Methods and Protocols aims to be helpful for all persons working with research and development of autoimmune laboratory diagnostics and for clinicians using autoantibody tests in daily work with patients.

This volume aims to provide clear and detailed methods to probe glutamate receptor function. Chapters in this book feature methods to study synaptic ultrastructure, receptor dynamics/receptor interactions and trafficking, cellular plasticity, receptor gene regulation, epigenetics, and clinical applications. In addition, it includes overview chapters that provide critical reviews and historical perspectives on particular techniques and fields of study. 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, Glutamate Receptors: Methods and Protocols covers well-established glutamate receptor techniques, as well as new and cutting-edge techniques designed so that students, researchers, and clinicians can gain an understanding of the issues surrounding each experimental technique and its potential applications.

This volume describes cell culture protocols for the maintenance, propagation, manipulation, and analysis of primary explanted cells from various mouse organ systems.

This third edition volume expands on the previous editions with new topics that cover drug discovery through translational bioinformatics, informatics, clinical research informatics, as well as clinical informatics.

This detailed second edition volume expands upon the prior edition by addressing newly emerged technologies as well as improved existing protocols in nucleic acid delivery via nanotechnology.

Chapters detail current status and perspectives of transgenic cotton, principle and methods for making transgenic cotton, creating gene knockout lines, foreign gene copy and expression in transgenic plants, improvements to cotton using transgenic technology, and monitoring the potential impact of transgenic cotton on environment.

New insights into modern medicine and systems biology are enabled by innovative protocols and advanced technologies in mass spectrometry-based proteomics. This volume details new pipelines, workflows, and ways to process data that allow for new frontiers in proteomics to be pushed forward.

Methods covering CRISPR gRNA design, CRISPR delivery, CRISPR activity quantification (indel quantification), and examples of applying CRISPR gene editing in human pluripotent stem cells, primary cells, gene therapy, and genetic screening are included.