Books in the Methods in Molecular Biology series

Divided into several sections, the book delves into animal-pathogen interaction, microbe-microbe interaction, as well as plant-pathogen interaction.

Methods for the successful delivery of these gene therapy vector systems to the retina are examined, as well as assays to test the efficacy in vitro in cell cultures, for gene augmentation and gene editing in vivo on rodents, pigs, and monkey retinas, and on human retinal explants as well as in human clinical studies.

This volume presents the basic concepts and practical experimental details investigators need to understand, develop, and successfully apply ELISA methodology. Includes numerous worked examples, detailed instructions, and extensive illustrations.

This open access volume gathers a variety of models, delivery systems, and approaches that can be used to assess RNA technology for exploiting antisense as a therapeutic intervention.

This volume details protocols on rationale design of therapeutic siRNA molecules and its encapsulation with smart vehicles to overcome the barriers to an effective administration 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 cutting-edge, Design and Delivery of SiRNA Therapeutics aims to ensure successful results in the further study of this vital field.

The aim of this volume is to provide a comprehensive description of methods and protocols useful for the further study of T-helper cells. Chapters guide readers through T-helper cell recovery, molecular study, signal transduction pathways, T-cell manipulation and, last but not least, ¿omic¿ 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. Authoritative and cutting-edge, T- Helper Cells: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field.

This detailed book brings together a new set of protocols to arm cell biologists with techniques that are currently being used in a number of well-established laboratories around the world. The contents represent the great strides made in the field of cell-cell communications with respect to the identification and characterization of key components of the communication apparatus, assembly and maintenance of the communications structures, and concomitantly their roles in not only tissue formation and maintenance but also regeneration and repair. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective chapters, 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, Stem Cell Renewal and Cell-Cell Communication: Methods and Protocols, Second Edition serves as an ideal guide for experts and newcomers alike seeking to increase our understanding of the crucial biological and physiological roles of cell-cell communications in tissue function and organismal integrity.

This volume explores the latest methods used to study various aspects of TET proteins and their biology. Chapters in this book are divided into five parts. Part One describes technologies aimed at detecting and quantifying DNA methylation turnover using massively parallel sequencing, ELISA, and mass spectrometry approaches. Part Two looks at data analyses protocols for distinguishing acting versus passive DNA demethylation and estimation of 5mC and 5hmC levels. Part Three deals with a new topic that takes advantage of modified CRISPR/Cas9 genome editing systems to target DNA demethylation activity to genomic loci of interest. Part Four discusses protocols that detail how to purify TET proteins and unravel their protein interactions, and Part Five looks at the assessment of TET protein function and activity in vivo and in vitro. 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, TET Proteins and DNA Demethylation: Methods and Protocols is a valuable resource that aims to help research scientists at all levels working in the fields of DNA demethylation dynamics.Chapters 3, 7 and 17 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume explores the techniques used to study human and animal pericytes. Chapters in this volume cover a range of topics such as pericyte dissociation from tissues, and analysis by immunohistochemistry and flow cytometry; avian chimaeras to investigate the embryonic development of cell lineage; pericyte purification by fluorescence activated cell sorting and ensuing culture; tissue turnover and regeneration; and the dynamic study of pericyte involvement in cancer dissemination, in situ, and in culture. 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, Pericytes: Methods and Protocols is a valuable resource that helps researchers define the biologic identity, functions, and potentials ofa cell lineage of increasing interest and therapeutic significance.

This volume provides methods and protocols on principle, production, molecular aspects, and sustainability of biofuel and biodiesel. Chapters are divided into four sections detailing biofuel production, biodiesel production, molecular genetics, biotechnology of biofuel and biodiesel production, economics, and sustainability of biofuel production. 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, Biofuels and Biodiesel aims to be useful for researchers, students, and enthusiasts in the field of biofuel and biodiesel.

This volume provides readers with a comprehensive look at the latest techniques used to identify and characterize PDZ-mediated interactions. Chapters cover topics such as promiscuity, multimodularity, regulation, and viral recognition by PDZ domains. 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, PDZ Mediated Interactions: Methods and Protocols is a valuable resource for all researchers interested in learning more about this developing field.

This second edition volume expands on the previous edition with updated chapters in model systems and new crops. The book contains protocols for plastid engineering in leaves, tissue culture cells, and the shoot apex of plants, as well as for marker excision from the plastid genome and engineering Rubisco, the key enzyme of photosynthesis. 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, Chloroplast Biotechnology: Methods and Protocols, Second Edition is a valuable resource for researchers who wish to enter the field, and for practitioners looking for insights of applications in agriculture, industrial biotechnology and healthcare.

This detailed book explores the fundamentals of myofibroblast biology in tissue repair, fibrosis, and tumors as well as providing cutting-edge laboratory methods used to investigate myofibroblast functions in physiological and pathological settings in vitro and in vivo, as written by leading academic scientists. Section I of this volume focuses on fundamental methods to study myofibroblast biology and covers topics such as methods for detecting myofibroblasts and senescent myofibroblasts in cell culture and histology, single cell RNA sequencing to identify myofibroblast subsets in fibrotic tissues, and functional assays to assess TGF-¿ activation, myofibroblast apoptosis, or matrix deposition and crosslinking. Section II discusses methods to investigate the mechanobiology of myofibroblasts in vitro, including the fabrication of functional hydrogels with tunable stiffness, the use of atomic force microscopy to characterize matrix and cellular stiffness, as well as molecular assays to assess fibroblast mechanotransduction pathways and durotaxis. Section III describes multiple animal models to investigate myofibroblast functions across organs in vivo as well as human organoid systems, precision tissue slices and decellularized 3D tissue scaffolds to assess myofibroblast functions in relevant human ex vivo models. 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 extensive, Myofibroblasts: Methods and Protocols is an essential collection for researchers delving into the processes and effects of these important cells.

This second edition provides new and updated chapters detailing recent advances in MYC research and current techniques. Chapters guide readers through protocols on how to express and purify MYC protein, X-ray crystallography, NMR, techniques to study how MYC is modified, apoptosis, senescence, proliferation, metabolic changes, translation, tumorigenesis,reprogramming, and clinical application of MYC studies.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, The Myc Gene: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

This volume provides a wide range of imaging protocols that can be tailored to specific organisms or cell-types. Chapters guide readers through fixed-cell, live-cell, phenotype screening, super-resolution, intravital imaging techniques, and fluorescence life-time imaging microscopy (FLIM). 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, Confocal Microscopy: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This volume presents the most useful laboratory protocols in strigolactones(SL) research. Chapters guide readers through wet-lab paths, issues around stability, protocols to evaluate SL activity, effects towards soil inhabitants such as parasitic plants, mycorrhizal and non-mycorrhizal fungi, nodulating bacteria, and protocols to assess effects on plant development are discussed. 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, Strigolactones: Methods and Protocols aims to deliver a clear-cut and standardized set of experimental protocols to a broad scientific community.

This second edition volume details the latest aspects of neural cells covering the practical and theoretical considerations of each techniques involved. Chapters guide readers through a general overview of the neuronal culturing principles, cell line models for neural cells, the isolation and propagation of primary cultures, stem cells, transfection and transduction of neural cultures, and other more advanced techniques. 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. Practical and easy to use, Neuronal Cell Culture: Methods and Protocols, Second Edition aims to be of interest to scientists at all levels studying cell culture models for neuroscientific studies.

This detailed book compiles a series of laboratory protocols covering the most important aspects of R-loop biology. Beginning with a range of methods allowing for the detection of DNA-RNA hybrids, as well as their purification and visualization by electron microscopy, the volume continues with methods based on the use of RNase H-derived tools to detect DNA-RNA hybrids in vitro and in vivo. Several protocols permit studying non-canonical RNA nucleotides in the R-loop context, as well as a number of specific protocols devoted to the investigation of R-loop topology and their functional roles in the biology of mitochondria and telomeres. Finally, a large block of chapters is dedicated to different methods allowing genome-wide mapping of DNA-RNA hybrids in various organisms. 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, R-Loops: Methods and Protocols serves as an ideal resource for those working on R-loop homeostasis but also to scientists studying such areas of molecular and cell biology as genome integrity, DNA replication and repair, chromatin remodeling, transcription, RNA processing, modification and export, as well as for researchers elucidating the molecular mechanisms of cancer and genetic diseases.

This volume details step-by-step instructions on biomedical literature mining protocols. Chapters guide readers through various topics such as, disease comorbidity, literature-based discovery, protocols to combine literature mining, machine learning for predicting biomedical discoveries, and uncovering unknown public knowledge by combining two pieces of information from different sets of PubMed articles. Additional chapters discuss the importance of data science to understand outbreaks such as COVID-19.   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, Biomedical Text Mining aims to be a useful practical guide to researches to help further their studies.