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This detailed book provides general protocols and technologies that plant biologists worldwide often utilize for the purpose of accelerating research progress in the field of plant transcription factors.

This volume provides the reader with step¿by¿step methods to study plant ARGONAUTE (AGO) functions. Chapters in Plant Argonaute Poteins: Methods and Protocols summarize the main biochemical, molecular and cytological methods to study AGO¿small RNA complexes and their interactions with target RNAs, AGO subcellular localization, AGO association with polysomes, AGO role in meiosis and DNA repair, methods for the identification, cloning and characterization of AGO genes in different plant species, and non¿radioactive protocols for small RNA detection. Finally, a series of chapters describing computational methods to study plant AGO function and evolution are provided. 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 Argonaute Proteins: Methods and Protocols aims to deliver the most complete and updated list of protocols to study plant AGO function.

This meticulous volume covers metabolism and drug-drug interactions during pregnancy, critical periods of developmental toxicology, in vivo and alternative methods to assess potential developmental toxicity for drugs and chemicals, and effects of chemicals on testes and mammary glands.

This volume provides key methods and protocols from laboratories engaged in germinal centers (GC) research with the expectation of stimulating further research, and to aid scientists in the study of GC biology and pathology.

This volume focuses on protein analysis covering a wide spectrum of the utility of mass spectrometry within neurobiological disciplines. The chapters in this book discuss label (iTRAQ, TMT, protein arrays) and label-free workflows (SWATH, MALDI Imaging, and label-free quantitation). Other chapters look at experimental strategies targeted to the identification and quantitation of specific lipids and post-translational modifications, as well as proteomic workflows that focus on characterization of subcellular proteomes. The last few chapters in the book describe various bioinformatics pipelines used to analyze the molecular data derived from high-throughput transcriptomic and proteomic experiments on brain tissue. The Neuromethods series offers chapters with key advice and procedure specifics to empower the readers to successfully achieve their own scientific and experimental goals.Thorough and comprehensive, Current Proteomic Approaches Applied to Brain Function is a valuable resource for graduate students and postdoctoral fellows interested in neuroproteomics, as well as researchers looking for further insight into the growing field of mass spectrometry in neuroscience.

Over the past decade, there has been an increase in powerful proteomics technologies that allow greater fundamental insights into the blood proteome. Further developments in informatic analyses, software developments, and computational tools are providing insights into large data sets, open-source data along with large-scale application of bioinformatics. Serum/Plasma Proteomics: Methods and Protocols, Second Edition is a comprehensive resource of protocols for areas, pre-analytical through to analytical, of plasma and serum proteomics. Divided into five convenient sections, covering fractionation strategies for in-depth blood proteome analysis, defined procedures for blood collection, handling and storage, detailed protocols for performing both antibody-based and non-antibody based quantitative assays, proteome analysis of blood cell compartments, circulating nanomebraneous vesicles and blood-related fluids, data management, statistical design, and bioinformatic challenges. 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, Serum/Plasma Proteomics: Methods and Protocols, Second Edition delivers a valuable foundation for the development and application of blood-based proteomics, and further incorporates blood cell components, including platelets, red blood cells, circulating extracellular vesicles/exosomes, and related biofluids.

This volume provides a collection of methodologies for basic research, clinical diagnosis, and treatment pertaining to food allergens, including food allergen production, purification, characterization, detection, quantification, and bioinformatics approaches to modern food allergen studies.

This volume provides a comprehensive collection of protocols on molecular diagnostics of bacteria that will suit the needs of molecular biologists, clinical laboratorians, and physician scientists alike.

The techniques described in Th9 Cells: Methods and Protocols study the role of Th9 cells in different inflammatory disease models, including allergic inflammation model, parasite model, tumor model, and EAE and IBD model.

This volume describes a broad spectrum of experimental approaches for investigating structure, function, and transport of neuronal mitochondria in health and disease. Most of these approaches were only recently developed and range from electron tomography-based 3D reconstruction of mitochondrial cristae to patch clamp recording from mitochondria in intact neurons. The chapters in this book cover topics such as mitochondrial proteomics, fluorescence lifetime imaging, respirometry and mitophagy, as well as optical approaches based on the use of genetically engineered fluorescent sensors for monitoring synaptic ATP and axonal ROS generation, mitochondrial Ca2+ cycling and pH changes, and mitochondrial dynamics and axonal trafficking in live neurons. Each chapter also discusses difficulties, tips, tricks, and precautions to take. 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, Techniques to Investigate Mitochondrial Function in Neurons is a valuable and useful resource for a broad range of investigators interested in the function of neuronal mitochondria in health and disease states.

While HIV-1 continues to be well-researched, this detailed volume draws attention to other members of the Retrovirus family, namely the Human T-lymphotropic Viruses (HTLVs), featuring the most updated technical information about HTLV determination and the methods to investigate their interaction with the host immune system and interfering pathogens. The contents include essential aspects of epidemiology and virus transmission, novel and robust methodologies for studying the effects of trans-activating regulatory HTLVs¿ proteins, the latest techniques for genotyping and gene expression analysis, as well as cellular phenotype and dynamics. 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, Human T-Lymphotropic Viruses: Methods and Protocols serves as an ideal guide to an area of study that is very much worthy of further research.

Since the identification of the first matrix metalloprotease (insterstitial collagenase or MMP-1) more than 20 closely related and evolutionarily conserved vertebrate MMPs have been discovered. Chapters in this volume detail methods and protocols on recent advances in the expression and purification of MMPs, biophysical methods such as X-ray crystallography, NMR spectroscopy and small angle X-ray scattering, computational and experimental methods for determining MMP specificity, methods for detecting MMP activity in vitro and in vivo, and methods for the development and characterization of MMP-based inhibitors as potential therapeutics for the treatment of various 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 practical, Matrix Metalloproteases: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This volume is essential for anyone struggling with the cellular heterogeneity field, and it connects this new area with the latest advances in innovative techniques combined with practical advice from international experts.

This detailed book provides a compilation of laboratory techniques and tests to assess the risks to embryo-fetal development from drug exposure during early developmental stages. After an introduction to teratogenicity testing, the contents explore both in vitro and in vivo techniques in the study of the teratogenic and fetotoxic effects of drugs, as well as numerous animal-based teratology methods. 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 comprehensive, Teratogenicity Testing: Methods and Protocols describes methods that integrate a battery of tests that can be performed in cells, organs, tissues, and animal models for evaluating toxicity and/or the safety of compounds in early developmental stages with the goal of estimating, preventing, or minimizing the teratogenic potential of drugs.

This volume provides practical experimental laboratory protocols for a wide range of steroid bioconversions. The chapters in this book cover topics such as bioconversions and chemical synthesis pathways; strain characterization; bioconversion from sterols to androstenedione and androstadienedione; steroid hydroxylations; biocatalysis; and downstream processes to purify steroid intermediates. 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, Microbial Steroids: Methods and Protocols is a valuable resource for laboratory and industrial professionals. It is also useful for graduate students studying biotechnology, microbiology, genetics, and molecular biology.

This second edition provides up-to-date and new chapters covering the most important widely-used techniques in plant stress tolerance, including cutting-edge strategies, in a manner that ensures effective results. Plant Stress Tolerance: Methods and Protocols, Second Edition guides readers through reviews on epigenetics, long noncoding RNAs, and the influence of microbiome on drought tolerance, the volume then continues with methods describing the identification of stress-responsive genes, proteins, posttranslational redox modifications, DNA methylation, histone modifications, measurement of glutathione, lipids, raffinose family oligosaccharides, polyamines, as well as quantification of abscisic acid, and the enzymes involved in ROS detoxification. 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 Stress Tolerance: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

This volume describes up-to-date techniques used in understanding the molecular biology of acute myeloid leukemia (AML) and addressing advances in diagnosis, classification, prognostication, and therapeutic strategies to potentially impact overall patient survival.

This volume details protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. 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, Genome Editing in Animals: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This vital book collects key methods that have supported advancements in the field of ebolavirus molecular biology given the pressing need for the advancement of techniques for diagnostics, the development of vaccines and antivirals, and for furthering our understanding of ebolavirus biology.

This volume presents protocols that analyze and explore gene regulatory networks (GRNs) at different levels in plants. This book is divided into two parts: Part I introduces different experimental techniques used to study genes and their regulatory interactions in plants. Part II highlights different computational approaches used for the integration of experimental data and bioinformatics-based predictions of regulatory interactions. This part of the book also provides information on essential database resources that grant access to gene-regulatory and molecular interactions in different plant genomes, with a specific focus on Arabidopsis thaliana. 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 cutting-edge, Plant Gene Regulatory Networks: Methods and Protocols is a valuable resource for scientists and researchers interested in expanding their knowledge of GRNs.

Chapters guide readers through comprehensive experimental approach for molecular investigations of plant receptor kinases, from the simplest methods for expression and purification of receptor domains to the most advanced methods aiming at understanding the dynamics of receptor complex formation and specificity of signaling pathways.

This thorough volume provides an in-depth introduction to and discussion of microRNAs (miRs) and their targets, miR functions, and computational techniques applied in miR research, thus serving the need for a comprehensive book focusing on miR target genes, miR regulation mechanisms, miR functions performed in various human diseases, and miR databases/knowledgebases. Without prior knowledge of the area of study, computational biologists, computer scientists, bioinformaticians, bench biologists, as well as clinical investigators will find it easy to follow the techniques in this collection. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detailed implementation advice that ensures successful results. Accessible and practical, Bioinformatics in MicroRNA Research functions as an ideal guide for researchers of all backgrounds to explore this vital area of study.

This volume addresses the various techniques and novel applications of mass spectrometry imaging (MSI) and its role as a discovery tool in the field of proteomics, lipidomics, and metabolomics. The chapters in this book demonstrate how MSI can be applied to many areas of research such as clinical pathology, translational medicine, toxicology, biomarkers and response studies, and potential incorporation of MSI into forensic workflows. 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. Innovative and comprehensive, Imaging Mass Spectrometry: Methods and Protocols is a valuable resource for research scientists and clinicians who are interested in further studies of MSI technologies.

This volume details protocols that cover the broad arsenal of techniques used to study a secretion system from A to Z. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as reporters to follow effector transport. 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, Bacterial Protein Secretion Systems: Methods and Protocol aims to provide techniques that are not restricted to the study of secretion systems but are also of specific interest for any researcher interested on multi-protein complexes of the bacterial cell envelope.

There are detailed methods on leading techniques for stem cell genetic engineering and intra-organ transplantation for research and therapeutic purposes, in vivo imaging of donor stem cells, and essential requirements for setting up a new stem cell laboratory.

This volume discusses protocols that aid in measuring different components in the renin-angiotensin-aldosterone system (RAAS). The book also looks at the methods used to assess angiotensin peptides and discerning the influence of RAAS components on different mammalian diseases. The chapters cover topics such as the use of fluorescent substrate to measure ACE2 activity in the mouse abdominal aorta, blood pressure monitoring using the radio telemetry method, and the analysis of angiotensin metabolism in the kidney using mass spectrometry. 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, The Renin-Angiotensin-Aldosterone Systems: Methods and Protocols is a valuable resource that provides scientists and researchers with the best approaches to examine RAAS.

This volume presents established bioinformatics tools and databases for function prediction of proteins. Reflecting the diversity of this active field in bioinformatics, the chapters in this book discuss a variety of tools and resources such as sequence-, structure-, systems-, and interaction-based function prediction methods, tools for functional analysis of metagenomics data, detecting moonlighting-proteins, sub-cellular localization prediction, and pathway and comparative genomics databases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, step-by-step instructions of how to use software and web resources, use cases, and tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Protein Function Prediction: Methods and Protocols is a valuable and practical guide for using bioinformatics tools for investigating protein function

This new edition provides updated and novel protocols of neuroproteomics methods that encompass both global-scale as well as targeted and specialized topics, which are timely additions for the molecular and phenotypic analysis of the central nervous system and CNS-related disorders.

This volume explores lipidomics through protocols that focus on areas of utility, techniques, and bioinformatics advancements. The protocols in this book cover topics such as isolation of specific membranes and specialized fractionation of subcellular compartments, and computational and functional analysis of lipid metabolizing enzymes. Each protocol also discusses methods of dealing with fractionated organelles¿cells to whole organisms. 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, Lipidomics: Methods and Protocols is a useful and valuable resource for biochemists, molecular biologists, and neuroscientists interested in neurology, ophthalmology and vision sciences, as well as mass spectrometrists with interest in disease discovery.

This thorough book covers the most recent proteomics techniques, databases, bioinformatics tools, and computational approaches that are used for the identification and functional annotation of proteins and their structure.