Bücher der Reihe Methods in Molecular Biology

This volume provides readers with a collection of new and classical methods, techniques, and applications used to address enduring questions about the structure and functions of the Golgi complex. The chapters in this volume cover diverse topics ranging from model systems; live and fixed cell imaging techniques; in vitro biochemical reconstitution systems; and specific methods developed to study Golgi formation, maintenance, and functions under physiological and pathological conditions. 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 authoritative, Golgi: Methods and Protocols is a valuable tool for researchers in the field who wish to explore new areas of Golgi biology and for new investigators interested in exploring Golgi structure and function.

This new edition collects diverse protocols compiled by experts from different areas of research as well as by biotech researchers developing novel technologies in the area of immunohistochemistry (IHC). After a few chapters to help orient novices entering the biomedical arena, the book includes methods chapters covering multiplex IHC including fluorescence and chromogenic techniques, combination of IHC with In Situ Hybridization (ISH), wavelet transform approach for organelle tracking, transcription factors in human stem cells, differentiation of mesenchymal stem cells, 3D imaging, phenotyping of glial cells in the human brain, tissue fixation, permeabilization, and cryo-preservation, as well as other topics. 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, Signal Transduction Immunohistochemistry: Methods and Protocols, Third Edition aims to arm novices and experts with vital protocols they can use either as-is or tailor them for specific experimental needs.

This volumes details methods focusing on technological innovation and recent advances in diabetes management. Chapters will guide readers through recent advances, beta-cell regeneration, non-invasive imaging of endogenous, transplanted islets theranostics, microRNA profiling of beta-cells and artificial intelligence, and deep learning algorithms in diabetes. 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, Type-1 Diabetes: Methods and Protocols is for a broad audience including basic researches, clinicians, and physician scientists whose major focus is in diabetes.

The volume presents a small selection of state-of-the-art approaches for studying transposable elements(TE). Chapters guide readers through HTS-based approaches, bioinformatic tools, methods to studyTE protein complexes, and the functional impact on the host. 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, Transposable Elements: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field. 

This volume provides up-to-date methods on single cell wet and bioinformatics protocols based on the researcher experiment requirements. Chapters detail basic analytical procedures, single-cell data QC, dimensionality reduction, clustering, cluster-specific features selection, RNA velocity, multi-modal data integration, and single cell RNA editing. 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, Single Cell Transcriptomics: Methods and Protocols aims to be a valuable resource for all researchers interested in learning more about this important and developing field.

This second edition provides state-of-the-art and novel methods on antibiotic isolation and purification, identification of antimicrobial killing mechanisms, as well as methods for the analysis and detection of microbial responses and adaptation strategies. Antibiotics: Methods and Protocols,  Second Edition, guides readers through updated and entirely new chapters on production and design, mode of action, and response and resistance. 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, Antibiotics: Methods and Protocols, Second Edition aims to inspire scientific work in the exciting field of antibiotic research.

This collection explores the use of pluripotent cells to generate early embryo-like structures in vitro, with the resultant 3D embryo-like structures in vitro phenocopying many of the developmental landmarks and 3D architectures seen normally in vivo. The protocols gathered herein cover diverse aspects of the topic and exemplify some of the key developments and improvements in the field. 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, Embryo Models In Vitro: Methods and Protocols serves as an ideal guide to these exciting and innovative advances in developmental biology.

This second edition provides new and updated methods detailing techniques and state-of-the-art approaches on the structure and function of plant endoplasmic reticulum (ER). Chapters guide readers through modern microscopy techniques, software protocols, purification, and analysis of ER membrane structure. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, The Plant Endoplasmic Reticulum: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

This detailed volume presents a variety of mouse models for cancer studies, from leukemia and lymphoma models to different types of subcutaneous and orthotopic models. Models for the healing process and the assessment of the immune response after local ablative therapies are also included, as well as imaging techniques that allow for the visualization of cancer at the cellular and tissue level. The book closes with a detailed description of necropsy, which is essential for obtaining good biomaterial for translational research. 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, Mouse Models of Cancer: Methods and Protocols serves as an ideal guide to these controlled and reproducible model experimental systems to study different aspects of cancer biology, including prevention, development, progression, and treatment.

This second edition provides updated and comprehensive methods on miRNA biogenesis and their role in the development and progression of various human diseases. Chapters detail miRNA biogenesis, isolating RNA, extracellular vesicles (EVs), circulating miRNAs, analyzing miRNA and miRDeep-P2, protocols for total RNA isolation from cells, cell-derived products, isolation and characterization of exosomes, serum, plasma specimens, and software tools. 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, MicroRNA Profiling: Methods and Protocols, Second Edition aims to provide comprehensive and accessible methods to undergraduate, graduate, and established scientist.

This volume details protocols emphasizing systems-level approaches that can be applied to genomic analyses. Chapters detail techniques for optimized application in in vivo systems, spatial, physiological, environmental contexts, imaging-based techniques, single-molecule approaches, CRISPR systems, new genomic approaches, and measurements of kinetics governing. 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, DNA-Protein Interactions: Methods and Protocols aims to present genome-wide techniques that will complement the biochemistry-based protocols to aid researchers in their studies. 

This fully revised new edition explores advances in the prevention and treatment of oral diseases. Beyond the updated chapters, the book delves into regenerative biology, gene editing and the use of CRISPR in oral biology, as well as histone acetylation and deacetylation methods, further reflecting advances in the application of molecular techniques to oral 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Oral Biology: Molecular Techniques and Applications, Third Edition serves as an ideal basic resource not only for new researchers but also for experienced scientists wishing to expand their research platform into new areas of this vital field.

 This second edition compiles state-of-art protocols to study the molecular mechanisms that determine proteome composition and contribute to maintaining protein homeostasis. Chapters describe methods that range from the in vitro analysis of enzyme cascades to real-time in vivo studies. 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, Plant Proteostasis: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field. 

This volume discusses the latest imaging and molecular techniques for studying microcephaly through neural progenitor proliferation, survival, and gene expression. All of the methods covered in this book use the mouse as a model system, and include cellular, metabolic, transcriptomic studies of neural progenitors, MR imaging studies of brain growth in 3-dimensions and developmentally-adapted studies of behavior in neonatal mice. 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, Microcephaly: Methods and Protocols is a valuable resource for all researchers who want to expand their knowledge and understanding of this important and developing field.

This second edition details new and updated methods on circular RNA. Chapter guide readers through circular RNA purification, in silico characterization, circRNA detection, sequence validation, quantification , techniques related to gain- and loss-of-function approaches, circular RNA synthesis, split ligation, engineering, nanoparticle packaging, RNA modifications on circular RNA biogenesis, RNA translation potential, and vaccines based on circular RNAs. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Circular RNAs, Second Edition aims to ensure successful results in the further study of this vital field.

This volume discusses the latest advancements in modern methodologies used to study liver carcinogenesis. The first half of this book describes pertinent preclinical models of hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA), established either through orthotopic induction of ectopic implantation. The second half of this book covers a diverse array of techniques applied to characterize the biochemical and cellular composition of hepatic malignancies that operate at the single-cell and histological levels. 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, Liver Carcinogenesis: Methods and Protocols, is a valuable resource for students and scientists who are interested in driving progress in the important field of liver cancer research.

This second edition explores standard laboratory protocols and methodology commonly used in basic and translational studies in the field of rheumatoid arthritis (RA) treatment. Chapters detail including basic RA models, evaluation of disease activity and immunological status, systemic drug delivery, and new research tools. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Rheumatoid Arthritis: Methods and Protocols, Second Edition aims to be a basic manual for clinical researchers who are just getting started in the field of intervention study.

 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 detailed volume explores techniques for the isolation, purification, and establishment of in vitro germ cell (GS) systems at different stages of development in a variety of organisms, including chickens, mice, rats, and humans. The book additionally describes cutting-edge analytic and informatic tools to study germ cell development at the single cell level and meiotic recombination. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Germ Cell Development: Methods and Protocols provides new insight into the understanding of basic biological aspects of germ cell biology, as well as the opportunity for in vitro manipulation of germ cells for toxicology studies and for genetic intervention.

This detailed volume examines classical and cutting-edge methods involving double-stranded RNA (dsRNA), specifically regarding isolation, visualization, characterization, production, and application. Many protocols, such as co-immunoprecipitation-based isolation of double-stranded RNA-associated protein complexes, identification of mycoviruses by dsRNA extraction, application of dsRNA for fungi disease management (Sclerotinia sclerotiorum and Botrytis cin), and production of double-stranded RNA in plants by plant viral vectors for gene silencing, can also be easily adapted for identification of viruses from other organisms, control of other pathogens, and fundamental research. 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 for troubleshooting and avoiding known pitfalls. Reliable and practical, Double-Stranded RNA: Methods and Protocols serves as an ideal reference book for students and researchers who work with dsRNA.

This volume explores the latest techniques used to study the human brain towards understanding Alzheimer¿s Disease and related neurodegenerative disorders. Contributed to by world-renowned experts, the chapters in this book are divided into five parts. Part One discusses human post-mortem brain preparations including single-cell isolation and use of specialized imaging. Part Two talks about neural cellular models using primary and human induced pluripotent stem cells to model aspects of the human brain. Part Three details nucleic acid analyses including transcriptomic and somatic genomic changes, and Part Four discusses lipid analyses via mass spectrometry. Lastly, Part Five covers protein analyses, particularly Aß and Tau. 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, Alzheimer¿s Disease: Methods and Protocols is a valuable tool for all researchers who want to expand their knowledge and understanding of this disease and other related dementias.

This volume explores the latest advancements in mucin research. The chapters in this book are organized into 8 parts and cover a wide range of topics such as mucin extraction, isolation, physicochemical property analysis, and experimental methods. The chapters also discuss the origins of mucins in jellyfish, feces, saliva and salivary glands, bronchi, stomach, and cervical tract; organic synthesis of peptides glycosylated at specific sites; analysis of mucin gene expression and methylation-specific electrophoresis of genes; imaging of mucin networks by AFM; and experimental methods using supported molecular matrix electrophoresis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, Mucins: Methods and Protocols is a valuable resource for researchers who want to obtain in-depth insight on how to more thoroughly conduct mucin research in their laboratories.

This detailed volume explores the production and purification of glycoproteins for therapeutic use or basic research, as well as the necessary in-depth understanding of cellular and acellular systems available for these purposes, their advantages and disadvantages, and considerations for choosing the most appropriate system for the desired application. Beginning with chapters on viral-encoded glycoproteins and those found in other pathogens, the book continues by examining the production of mammalian glycoproteins, the analysis of glycoprotein content, cell-free synthesis of glycoproteins and considerations for production and purification of glycoproteins. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to the 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, Recombinant Glycoproteins: Methods and Protocols provides a wide range of guidelines for studying these vitally important proteins.

This detailed volume explores the most recent methodologies and applications of stable isotope labeling by amino acids in cell culture (SILAC). With the development of new technologies in the field of mass spectrometry and bioinformatics, SILAC-based methods have become the first tool of choice in quantitative proteomics either on its own or in combination with other more recently developed approaches, which is elucidated in the chapters of this book. 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, SILAC: Methods and Protocols serves as an ideal guide for researchers working in the area of functional proteomics and other aspects of protein science.

This detailed volume provides methods to guide assay development, procedures designed to investigate the chemokine and glycosaminoglycan (GAG) networks, as well as their interactions, in a wide range of organs and tissues in disease and in health. The initial chapters in this book present in vivo models used to examine the roles of chemokines and GAGs in normal physiology and in the pathophysiology of disease. The book then explores present cell- and tissue-based in vitro assays to examine chemokine:GAG interactions. Finally, analytic approaches are presented that provide assays for measuring GAGs, chemokines, and cellular responses. 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, Chemokine-Glycosaminoglycan Interactions: Methods and Protocols serves as an ideal guide for researchers seeking to analyze chemokine and GAG functions, interactions, and molecular mechanisms in vivo and in vitro.

The second edition of this volume provides a comprehensive update of this key method on gel-based proteomics. Chapters present an introduction into the development of methods on principles of differential protein labeling and two-dimensional gel electrophoresis, techniques on optimized proteomic workflows using advanced mass spectrometry for protein identification, and the application of those methods in basic biological research, pathobiology and applied biomarker discovery. 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, Difference Gel Electrophoresis: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field. 

This second edition volume expands on the previous edition with updated methodologies and practical tips to overcome obstacles associated with experimentation pertaining to chemistry, biology, physiology, pathology, medical and dental sciences, and pharmacology of CCN proteins. The chapters in this book cover topics such as CCN4 immunofluorescence for tissue microarray; utilizing public molecular biological databases for CCN family research; the effects of CCN4 on pancreatic beta cell proliferation; gene expression analysis of CCNs; novel cell biological assays for measuring bone remodeling activities of CCN proteins; and the function of CCN2 in tubular epithelium cells with a focus on renal fibrogenesis. 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, CCN Proteins: Methods and Protocols, Second Edition is a valuable resource for experienced CCN researchers looking for new approaches and novice researchers just starting out in the field of CCN research.

This detailed book presents a comprehensive collection of state-of-the-art protocols on muscular dystrophy therapeutics, covering therapeutics using antisense oligonucleotides, gene replacement, genome editing, small molecules, stem cells, and antibodies. Written by leaders in the field, the volume explores techniques that are currently in use and are starting an exciting therapeutic revolution in muscular dystrophy. As a part of the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Muscular Dystrophy Therapeutics: Methods and Protocols serves as an ideal resource to inspire readers and provide tips, strategies, and advice to develop new therapeutic technologies for this group of diseases.

This second edition volume provides new and updated chapters detailing simple and accessible experiment protocols to explore thymus biology. Chapters are divided into three parts presenting short reviews, analysis strategies, protocols for cell preparation, flow cytometry analyses, Innate Lymphoid Cells (ILC), mouse T-cell development, antigen receptor-less cousins of T cells, bone marrow chimeras, thymic stroma, and multiple aspects of thymocyte biology. 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, T-Cell Development: Methods and Protocols, Second Edition aims to be a useful practical guide to help readers overcome obstacles associated with experimental approaches of T-cell development.