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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 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 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 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 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 discusses protocols that cover genetic manipulation of Chinese hamster ovary (CHO) cells for recombinant protein production, and protocols for the characterization of CHO cells using 'omic approaches.

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 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 details methods for the analyses of specific lipid classes and lipidomics analyses of cells such as lymphocytes and oocytes.

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 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.

The chapters in this book discuss the use of fluorescent metal probes, synchrotron-based X-ray microscopy, ICP-MS, laser ablation-ICP-MS, laser-based tissue microdissection, MRI image analysis, fractionation of cell tissue samples for metal analysis, and metal treatment of cells.

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 detailed volume focuses on population epigenetics, which is of increasing interest to policy makers searching for explanations for complex epidemiological observations and conceptual models on which to base interventions.

This volume provides comprehensive and detailed technical protocols on current biosensor and biodetection technologies and examples of their applications and capabilities. Chapters in Biosensors and Biodetection: Methods and Protocols Volume 2, Electrochemical, Bioelectronic, Piezoelectric, Cellular and Molecular Biosensors, Second Edition focus on electrochemical biosensors including amperometric, impedance and voltammetric sensors, bioelectronic, piezoelectric, cellular, and molecular biosensors. 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, tips on troubleshooting and avoiding known pitfalls, and step-by-step, readily reproducible laboratory protocols.Authoritative and practical, Biosensors and Biodetection: Methods and Protocols Volume 2: Electrochemical, Bioelectronic, Piezoelectric, Cellular and Molecular Biosensors, Second Edition offers descriptions of major technologies by leading experts in the field in extensive technical detail. The aim of the book is to make biosensors more accessible and understandable to engineers, students, medical professionals, molecular biologists, chemical, and physical science researchers developing biosensor technologies, allowing readers to both understand the technology and to construct similar devices.

This third edition volume expands on the previous editions with new and improved assays that ask essential questions of telomere regulation in diverse model systems. The chapters in this book discuss genetic, proteomic, genomic, biochemical, and molecular approaches that provide insight into the complex protein interaction networks active in the telomere chromatin. This book also details information regarding telomere dynamics in response to stress or stimuli. 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, Telomeres and Telomerase: Methods and Protocols, Third Edition is a valuable and informative resource for anyone who is interested in the developing field of telomeres and telomerase.

This volume is a wide-ranging tool for studying protein-carbohydrate interactions that extend from traditional biochemical methods to state-of-the-art techniques.

This volume provides state-of-the-art techniques for studying various aspects of cholesterol homeostasis, including its uptake, synthesis and efflux from the cell, as well as its trafficking within the cell.

This volume provides all the essential protocols that are currently used to study the immune synapse. Chapters in The Immune Synapse: Methods and Protocols cover methods for the study of the dynamics of immune synapse assembly, traffic at the immune synapse, new high resolution imaging, biophysical and computational methods for the study of the immune synapse, effector immune synapses, B cell, NK and mast cell immune synapses, and immune interactions in vivo. 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, The Immune Synapse: Methods and Protocols will be of interest to immunologists and, at a more general level, to cell biologists, biophysicists and computational biologists.

This detailed volume provides a toolbox for designing constructs, tackling expression and solubility issues, handling membrane proteins and protein complexes, and exploring innovative engineering of E.

This volume is a collection of miRNA detection and target identification protocols, detailing new developments in the traditional detection approaches such as northern blot, quantitative real-time PCR, array, next generation sequencing, and in situ hybridization.