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This volume provides a comprehensive and technical presentation of numerous aspects of reproductive cell tissue cryopreservation, and presents readers with current procedures and detailed discussions of novel techniques and the latest innovations. The chapters in this book are divided into five parts and cover subjects such as: immature oocyte cryopreservation, human sperm vitrification and slow-freezing, directional freezing of ovarian tissue, automated vitrification systems, and detailed protocols on popular and commercially available cryopreservation/vitrification systems. 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, Cryopreservation of Mammalian Gametes and Embryos: Methods and Protocols includes chapters written by leading experts in the field and is a valuable resource for anyone interested in the field of cryopreservation.
This volume focuses and describes tools, assays, and techniques used to enhance the understanding of the role of auxins and cytokinins. The chapters in this book cover topics such as: microbial manipulation of auxin and cytokinins in plants; interplay between auxin and cytokinin and its impact on MAPK; H2O2 production in Arabidopsis leaves; crosstalk between jasmonate and auxin in plant stress responses of roots; and high-throughput protoplast trans-activation (PTA) screening. 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, Auxins and Cytokinins in Plant Biology: Methods and Protocols is a valuable resource to plant scientists, graduate and under graduate students in addressing their biological questions relevant to the functional implications of auxin and cytokinins.
This detailed volume compiles state-of-the-art methodologies for the study of brassinosteroid hormones, contributed by recognized researchers in the field, in order to bring together different experimental and theoretical biology techniques for addressing the questions of how brassinosteroids function in Arabidopsis and other agriculturally valuable species, such as rice and sorghum. 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, Brassinosteroids: Methods and Protocols assesses a wide range of appealing methodologies for experts as well as researchers with a general interest in hormone signaling in plants.
This volume presents a historical perspective on Morpholinos, an overview of good Morpholino practices, techniques for controlling Morpholino activity with light, techniques for modulating microRNA activity in zebrafish embryos, probing genes during fin regeneration, methods for determining to structure of gene networks during development, electroporation, bacterial knockdowns, pretargeting, diagnostic applications of Morpholinos, techniques for delivering Morpholinos in utero to developing embryos, and methods offering rapid, hands-free, label-free and inexpensive assays of Morpholino concentrations in biological extracts. 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.Morpholino Oligomers: Methods and Protocols aims to provide a diverse application of Morpholinos along with protocols that will assist new labs in moving the frontier.
This volume presents a collection of protocols that describe methodologies to study thermogenic fat biology from various angles. This book is divided into 2 parts. Part 1 focuses on establishing in vitro culture systems. The chapters in this section introduce techniques on how to isolate, culture, and differentiate primary fat cells from both laboratory mice and humans. This part also presents flow cytometry methods to isolate various subpopulations of precursors within the stromal vascular fraction of the adipose tissue, which contains both preadipocytes and immune cells. Part 2 introduces multiple means to genetically manipulate and evaluate brown and beige fat in vivo. The chapters in this section explore methods on bioenergetics analyses both in vitro and in vivo. They also cover how to evaluate thermogenic fat contents and activity in humans, how to culture these cells though interdisciplinary approaches, and how to use thermogenic fat cell lines to carry out drug screens. 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, Thermogenic Fat: Methods and Protocols is a valuable resource for both experts and novices who are interested in learning assays and investigating brown and beige fat functions.
This volume addresses up-to-date light microscopy approaches and toolsets offered for live- or fixed-cell observations. The imaging strategies discussed in this book include confocal laser scanning and spinning disk confocal microscopy, FRET, FRAP, and laser microsurgery experiments. Chapters also describe the use of these imaging methodologies to study properties of a multitude of biomolecular targets in a broad range of model systems ranging from bacteria over tissue to whole animal imaging. Light Microscopy: Methods and Protocols puts special focus on system instrumentation parameters and sophisticated labeling and detection 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.Cutting-edge and thorough, Light Microscopy: Methods and Protocols offers the novice user with straightforward strategies to address biological questions, while providing the experienced researcher with the latest applications that can be useful in routine practices. This book also serves as a useful teaching manual in practical courses on light microscopy.
This volume provides a comprehensive collection of current methods and protocols to study posttranscriptional base modifications in RNA with special focus on methylation.
This volume provides leading-edge protocols in the study of the molecular and cellular biology of muscle stem cells.
This volume introduces bioinformatics research methods for proteins, with special focus on protein post-translational modifications (PTMs) and networks.
Inflammation: Methods and Protocols is divided into four sections: the first three sections describe protocols investigating immune-mediated inflammatory disease models affecting barrier organs to the environment;
This volume covers an array of techniques available for studying SH2 domains and phosphotyrosine signaling. The book is divided into six parts: Part I outlines the history of SH2, technology development, and cell signaling; Part II focuses on computational approaches and tools used for identification, classification, and predictions of SH2 domain binding partners; Part III details various ways to prepare the SH2 domains as experimental reagents; Part IV presents methods for structural analysis and conventional binding assays using SH2 domains; Part V describes high-throughput and proteomics approaches to aid in analyzing SH2-mediated interactions; and Part VI covers applications for SH2 domain to functional and imaging analyzes. 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, SH2 Domains: Methods and Protocols is a valuable resource for computational biologists, biochemists, structural biologists, cell biologists, pathologists, and people interested in SH2 domains and phosphotyrosine signaling. Researchers who are investigating how protein interaction domain mediate specificity in signaling systems may also find this book informative.
This volume provides basic and advanced laboratory protocols for the non-specialist and the experienced researcher. Chapters are divided into three parts covering synthesis and characterization of AMPs, studying the interaction of AMPs with model systems or bacteria, and assaying selected biological activities of AMPs. 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, Antimicrobial Peptides: Methods and Protocols aims to ensure successful results in the further study of this vital field.
This volume aims to provide protocols on a wide range of biochemical methods, analytical approaches, and bioinformatics tools developed to analyze the proteome.
This volume aims to provide a new perspective on the broader usage of Hidden Markov Models (HMMs) in biology.
This volume provides detailed techniques used for the study and characterization of the plant vascular system, with a central focus on the xylem tissue. analysis of xylem development, xylem characterization though imaging techniques, and analysis of the xylem composition.
This volume illustrates how microfluidic approaches can meet the requirement of clinical diagnosis based on molecular or cellular biomarkers. Microchip Diagnostics: Methods and Protocols is divided into four sections describing the business aspects of the microfluidic-based systems for diagnosis, demonstrating how versatile microfluidics can be regarding to protein bioassay integration, presenting microfluidic approaches for nucleic analysis based on mono or diphasic format, and highlighting recent contributions. 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.
This volume serves as a reference for the dissemination of advances made in the study of Hepatitis B Virus (HBV). Hepatitis B Virus: Methods and Protocols details protocols and techniques ranging from cell culture studies to in vivo and clinical immunology. The chapters in this book discuss treatments of in vitro infection systems, analysis and quantification of cccDNA and its mutations; in vitro polymerase activity assays; cellular trafficking of core proteins; intracellular calcium metabolism; detection, cloning, and sequencing of HBV markers; and new strategies aimed at exploiting new mechanisms for drug discovery. The book also covers classical methods for resolution of extracellular viral particles by native gel electrophoresis, and methods for detecting HBV antigens in drug 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.Cutting-edge and comprehensive, Hepatitis B Virus: Methods and Protocols is a valuable tool for researchers to use toward their advanced studies in HBV.
This detailed volume presents protocols for advancing the utility of nanotechnology in cancer research toward improving our understanding of cancer biology, prevention, diagnosis, and therapy. There are continuous new discoveries in the field of nanotechnology, thus creating new imaging systems or therapies, and this book focuses on how to employ certain discoveries for studying cancer by presenting principles along with techniques to allow for the transformation of any new discoveries in the field into cancer-studying tools with the hope of bringing in the involvement of biomedical scientists who can enhance the speed of discoveries toward cancer diagnosis and therapy. 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 motivating, Cancer Nanotechnology: Methods and Protocols serves as an ideal resource for biomedical scientists interested in the potential of this field as well as for physical scientists and engineers interested in employing nanotechnology in cancer diagnosis and therapy.
This volume describes a variety of protocols that will allow the readers to study different aspects of transcriptional and posttranscriptional gene expression regulation in eukaryotic cells.
This volume aims to present a representative cross-section of modern experimental approaches relevant to Plant Hormone Biology, ranging from relatively simple physiological to highly sophisticated methods. Chapters describe physiological, developmental, microscopy-based techniques, measure hormone contents, and heterologous systems. 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, Plant Hormones:Methods and Protocols, Third Edition aims to provide researchers with useful methods to advance their research.
This volume presents classical approaches to in vivo neuropharmacology and neurophysiology, such as c-fos, electrochemistry, microdialysis microstimulation, and push-up superfusion. It also explores exciting new methods for behavioral analysis, and techniques based on optogenetics and non-invasive magnetic resonance imaging. The chapters of this book cover topics such as principles of stereotaxy, pharmaco-based fMRI and neurophysiology in humans and non-human primates, electrical nerve stimulation and central microstimulation, involvement of neurotransmitters in mnemonic processes, the impact of cannabinoids on motor activity, as well as the involvement of nitric oxide in neurotoxicity produced by psychostimulant drugs. 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 own laboratory.Cutting-edge and practical, In Vivo Neuropharmacology and Neurophysiology is a valuable resource for experienced and less experienced investigators of brain function and brain disorders.
This volume explores a broad range of different genotyping techniques. Genotyping: Methods and Protocols consists of chapters that cover numerous topics such as: an overview of multiplexed microsatellite analysis; High Resolution Melt analysis and TaqMan-based assays; in situ analysis of variants in single RNA molecules; the MassARRAY system and Molecular Inversion Probes; Pulsed Field Gel Electrophoresis, Paralogue Ratio Test, and Multiplex Ligation-Dependent Probe Amplification; long-range PCR combined with PacBio sequencing; Targeted Locus Amplification; Multilocus Sequence Typing and rapid SNP detection with pyrosequencing; and genotyping-by-sequencing for plant analysis. Finally, the volume concludes with a summary of pertinent points to describe genetic variation. 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 practical, Genotyping: Methods and Protocols is a valuable resource for anyone interested in learning more about the diverse field of genotyping.
This collection seeks to elucidate the practical methods necessary for successful adjuvant development, with a particular focus on the synthesis, formulation, manufacturing, and characterization aspects involved. Beginning with an overview and a case study, the book then delves into in silico design, chemical synthesis, biosynthesis, and/or purification from natural raw materials of specific adjuvant molecules, adjuvant formulation approaches, the analytical characterization of adjuvant formulations and adjuvant-containing vaccines, as well as the biological characterization of vaccine adjuvant activity, including in vitro and in vivo approaches, to measure innate and adaptive immune responses. Written in the highly successful Methods in Molecular Biology 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 authoritative, Vaccine Adjuvants: Methods and Protocols aims to facilitate vaccine adjuvant product development efforts, making them more accessible, manufacturable, and better characterized.
Now poised to evaluate the potential in modulating Wnt signaling for therapeutic agendas in cancer, wound healing, and degenerative disease, this book collects protocols from an exciting area of study.
Practical and cutting-edge, Plant Synthetic Promoters: Methods and Protocols covers all steps required from the identification of cis-regulatory elements, over synthetic promoter design, to the experimental analysis of synthetic promoter function.
This thorough volume explores predicting one-dimensional functional properties, functional sites in particular, from protein sequences, an area which is getting more and more attention. Beginning with secondary structure prediction based on sequence only, the book continues by exploring secondary structure prediction based on evolution information, prediction of solvent accessible surface areas and backbone torsion angles, model building, global structural properties, functional properties, as well as visualizing interior and protruding regions in proteins. Written for the highly successful Methods in Molecular Biology series, the chapters include the kind of detail and implementation advice to ensure success in the laboratory. Practical and authoritative, Prediction of Protein Secondary Structure serves as a vital guide to numerous state-of-the-art techniques that are useful for computational and experimental biologists.
High-throughput RNAi screening remains one of the most widely used technologies to perform target identification and validation studies in an unbiased manner. These assays are equally important for research and development across academic, biotech, and pharmaceutical industries. The success of these screening efforts is dependent on robust methodologies to perform these screens. In High-Throughput RNAi Screening: Methods and Protocols, expert researchers in the field share protocols and methods for performing high-throughput RNAi (HT-RNAi) screens. These include the use of various RNAi platforms and delivery methods in mammalian and non-mammalian systems, whole organism and cell models, and various applications, such as drug sensitizer identification. Finally, the book examines the latest advancements in the fields of assay development, library screening, data analysis, and hit selection. 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, High-Throughput RNAi Screening: Methods and Protocols provides a comprehensive source of protocols and other necessary information to make robust and successful assays possible for all who wish to apply HT-RNAi in their research.
This volume presents authoritative and cutting-edge methods and protocols focusing on three tool boxes covering the increasingly diverse methodologies used to image selected proteins and to investigate their function by light and electron microscopy.
This second edition provides 21 new chapters on methods used in laboratories for investigating the physiology and molecular genetics of the pathogen Clostridium difficile.
This volume provides a unique forum to review cell microencapsulation in a broad sense by exploring various cell types that have been encapsulated for different purposes, different approaches and devices used for microencapsulation, the biomaterials used in cell microencapsulation, the challenges to the technology, and the current status of its application in different clinical situations. This book is divided in five sections: Section I is an introductory part that discusses historical developments of the technology and its current challenges, as well as the various applications of cell microencapsulation; Section II discusses the main approaches and devices currently used in cell microencapsulation; Section III presents an overview of the various polymeric materials currently in use for cell microencapsulation and the enabling technologies to either monitor or enhance encapsulated cell function; Section IV gives specific examples of the methods used to encapsulate various cell types; and Section V provides an overview of the different clinical situations in which cell microencapsulation has been applied. 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 practical, Cell Microencapsulation: Methods and Protocols is a valuable reference for researchers, engineers, clinicians, and other healthcare professionals, as well as food technologists who will find detailed descriptions of methods for the microencapsulation of specific cell types and their current of potential clinical and industrial applications. This volume also includes detailed information about the design and manufacture of different devices including large-scale production devices for use in cell microencapsulation.
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