Bøger udgivet af Humana Press Inc.

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.

The second edition of this volume provides up-to-date methods on the main methodological aspects of functional MRI (fMRI), applying fMRI to the study of central nervous system, and future evolutions of fMRI techniques.

This volume provides a comprehensive overview of the experimental and computational methodologies used to study the function of long non-coding RNA (ncRNAs) expressed from enhancers. Chapter detail both wet-lab and dry-lab techniques and annotating long ncRNAs and exploring transcription by assessing where transcription starts and generally how it occurs.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, Enhancer RNAs: Methods and Protocols aims to ensure successful results in this rapidly developing field.

This second edition provides updated and new chapters on selected genetic, molecular, biochemical, and cell biological techniques. Chapter's guide readers through methods and principles on primordial germ cells and germline stem cells, however many of these principles can be applied to different types of adult stem cells.

This volume discusses experimental brain injury models thatcontain valuable information carefully chosen to widen the researchers¿ horizonabout neurotrauma. Injury Models of Central Nervous System: Methods and Protocols contains relevant experimental design approaches that have beenadapted and made ready for application in laboratory settings. For easiernavigation, the chapters are categorized into 6 parts: Introduction, GeneralConsideration in Using Animal Laboratory in CNS Injury Research, Classical TBIModels and Their Link with Pathophysiological Features of CBS Injury ¿ Models,Special Topics in CNS Trauma: Comorbid Conditions in CNS Injury, OutcomeMeasures in Brain Injury Models, and Future Directions. Written in the highly successful Methods inMolecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessary materials and reagents, step-by-step,readily reproducible laboratory protocols, and tips on troubleshooting andavoiding known pitfalls.Practical and thorough, Injury Models of Central Nervous Systems: Methods and Protocols, is a very useful reference towards theprogress of this discipline.

This volume highlights the molecular and cellular methods used in studying Chronic Myeloid Leukimia (CML) pathogenesis and stem cell biology. 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, Chronic Myeloid Leukemia: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This book covers core and emerging in vitro and in vivo protocols used to study how various components of the tumor microenvironment are established and subsequently interact with tumor cells to facilitate carcinogenesis.

This volume presents relevantbackground information to understanding the molecular basis governingunconventional protein secretion (UPS), and in particular explores the latesttechniques and protocols that have been successfully applied for the study ofthis topic. Detailed chapters include an overview of conventional andunconventional secretory pathways along with multidisciplinary approaches andmethods used for UPS analysis in different organisms. Written in the highly successful Methods inMolecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessary materials and reagents, step-by-step,readily reproducible laboratory protocols, and tips on troubleshooting andavoiding known pitfalls. Authoritative and cutting-edge, UnconventionalProtein Secretion: Methods and Protocols will be usefulfor all interested in the secretory pathway field as well as applications incell biology, cell development, biomedical research, and healthcare.

This volume is the result of an explosion ofmolecular-based research on Cilia, which began with the discovery of theuniversality of intraflagellar transport (IFT) and ciliary genomics/proteomics.The chapters in this book cover topics such as: high resolution imaging andfunctional characterization of sensory and primary cilia in mammalian cells andzebrafish, methods to study ciliary-mediated chemoresponse in Paramecium,and methods to study centrosomes and cilia in C. elegans and Drosophila.Written in the highly successful Methods in Molecular Biology series format,chapters include introduction to their respective topics, lists of thenecessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls.Practical and cutting-edge, Cilia:Methods and Protocols is broad and covers motile, sensory, and primarycells. It is a valuable resource to anyone interested in entering the field ofciliary biology using model organisms, including flagellate algae, ciliates,planaria, nematodes, insects, zebrafish, and mammalian cells.

This volume provides an up-to-date compilationof current methodological approaches utilized for the exploration of nucleolarstructure and function. Chapters covera diversity of protocols that include imaging of the nucleolus, analysis ofribosomal RNA transcription and processing, and genomics and proteomics of thenucleolus.

This volume presents methods and techniques to studyoogenesis in a broad range of organisms, from plants to mammals. Oogenesis: Methods and Protocols guidesreaders through protocols on models ofdevelopmental biology, oogenesis in plants, worms, fruit flies,mosquitos, butterflies, starfish, zebrafish, frog, chicken and mouse. Written in the highly successful Methodsin Molecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessary materials and reagents, step-by-step,readily reproducible laboratory protocols, and tips on troubleshooting andavoiding known pitfalls.Authoritative and cutting-edge, Oogenesis:Methods and Protocols aims to ensure successful results in the furtherstudy of this vital field.

Thissecond edition details new emerging areas of zebrafish research focusing on geneticsand genomics, techniques for developing and analyzing zebrafish disease models,and methods for neuroscience. Zebrafish:Methods and Protocols, Second Edition guides readers through methods for mutagenesisand genome editing in zebrafish, applicationsof GFP-expressing transgenic fish, techniques for cancer models, imaging of infectionand host-pathogen interactions, metabolism and transport of lipids, and the structure and function of neural circuitsand their role in generating behavior. Written in the highlysuccessful Methods in Molecular Biology series format, chapters includeintroductions to their respective topics, lists of the necessary materials andreagents, step-by-step, readily reproducible laboratory protocols, and tips ontroubleshooting and avoiding known pitfalls. Authoritative and practical Zebrafish: Methods and Protocols, Second Edition is a usefulcomplement to the first book for new and experienced zebrafish researcheralike.

This volume provides different methodologies for all classes of histone deacetylases, which includes detailed procedures on Class I and II histone deacetylase inhibitors, SIRT inhibitors, and bromodomain inhibitors.

This book provides coverage, methodology, and laboratory protocols on the more essential aspects of protein tyrosine phosphatase (PTP) function and regulation, including the use of standardized in vitro functional assays, suitable cell systems, and animal and microorganism models.

This volume highlights the role of proteostasis in human health and associated disease model systems, reflecting its rising importance which has led to the development of new technologies to obtain insight into underling protein mechanistic events.

This volume aims to outline the current status of the Mesenchymal Stem Cells(MSC) field in regenerative medicine and to propose clear and reproducible protocols to better define the identity, function and use of these cells that are today, more than ever, ¿under the spotlight¿. Mesenchymal Stem Cells: Methods and Protocols, Second Edition is organized into four sections. The first guides the reader through a series of state-of-the-art reviews summarizing the use of MSC for the treatment of various diseases. The other three sections are a collection of methodological chapters covering several aspects: isolation and characterization of MSC; expansion of MSC for clinical use; production and characterization of the MSC secretome. Written in the highly successful Methods in Molecular Biology series format, the method chapters include introductions to their respective topics, complete lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting which will help the researcher to avoid known pitfalls.Authoritative and cutting-edge, Mesenchymal Stem Cells: Methods and Protocols, Second Edition, aims to ensure successful results in the further study of this vital field.

This volume covers arange of methods used in plant cytogenetics, beginning with basic analysis ofchromosomes and visualizing gene locations, to manipulating and dissectingchromosomes, and then focusing on less understood features of chromosomes suchas recombination initiation sites and epigenomic marks.

Thisvolume encompasses all major methodologies to interrogate endocannabinoidsystems (ECS) and endocannabinoids (eCBs) signaling. With increasing interesttowards the manifold activities of eCBs, this book discusses the chemical,biochemical, and molecular biological assays, and activity of distinct elementsof the ECS. These include membrane, nuclear receptors, biosynthetic andhydrolytic enzymes, and membrane transporters and oxidative enzymes. Written inthe highly successful Methods in Molecular Biology series format,chapters include introductions to their respective topics, lists of thenecessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls.Timely and cutting edge, Endocannabinoid Signaling: Methods and Protocolsis a valuable resource and will help chemists, drug designers, biochemists,molecular biologists, cell biologists, pharmacologists, and (electro)physiologists navigate the mare magnum of endocannabinoid research.

This text provides a comprehensive, state-of-the-art review of the application of image analysis focusing on the techniques which can be used in every biology and medical laboratory to automate procedures of cell analysis and to create statistics very useful for a comprehension of cell growth dynamics and the effects of drugs on them.

This volume provides researchers with up-to-date protocols and takes aclose look at current research and promising applications.

methods used in both clinical and research laboratories for testing genetic alterations (Genetic Detection of Breast Cancer) and (Isolation of Breast Cancer Cells); and methods used for mimicking human breast cancer in a living organism (In Vivo Experimental Models for Breast Cancer).

This volume provides a detailed step-by-step description of protocols for the establishment of in vitro cultures of important medicinal plants, their mass multiplication in controlled environment, and step-wise secondary metabolite analysis.

The Sample Preparation Techniques for Environmental, Plant, and Animal Samples handbook is a collection of best practices, recipes and theoretical information aimed at anyone who works with any type of molecular biology, proteomics, or metabolomics research involving diffi cult and tough-to-process samples, and thus is exposed to the seemingly unbreakable bottleneck of sample preparation. Th is book is most useful to researchers preparing nucleic acids and proteins from environmental (e.g., soil, marine, and wastewater, feces) and tough microbiological (e.g., spores, yeasts, gram positive bacteria) samples, as well as solid tissue samples from plants and animals. This book is the first comprehensive piece of literature dealing with applications of bead beating technology and other types of mechanical homogenization sample preparation.

This volume explores the role free radicals and antioxidants within the development of vascular disease, examining fundamental research and translating preclinical knowledge to clinical trials. The expertly authored chapters describe the relationship of oxidative stress to atherosclerosis and the cardiovascular system, exploring its role in cardiac fibrosis, renovascular disease, hypertension, and regulation of blood pressure and cerebral vascular tone. The concluding chapter discusses the current state of clinical research, contextualizing clinical trials within the existing theoretical framework and analyzing attempts to preserve oxidant stress under various conditions. With its concise and authoritative analysis of pre-clinical research and clinical results, Studies in Atherosclerosis - part of the bestselling Oxidative Stress in Basic Research and Clinical Practice series - is essential for researchers and clinicians focusing in cardiology, nephrology, or oxidative stress.

This volume provides detailed molecular analysis of the associated pathways and cell types involved in Celiac Disease, and embraces new technologies such as next generation sequencing. The book is divided into three parts: Part I gives a background overview with a number of reviews to describe the history and nature of the disease, its diagnosis, the role of animal models, and study designs for investigating genetic susceptibility. Part II describes molecular techniques, including tissue culture, isolation and cloning of relevant cell types, high content analysis of biopsies, and HLA genotyping. The final part of the book begins with an outline of scripting for data management, focusing on tools that are freely available to researchers who wish to explore them. It also describes analysis pipelines for bioinformatics prediction of antigenicity, quality control and analysis of GWAS data, and transcriptome analysis by next generation sequencing. 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 thorough, Celiac Disease: Methods and Protocols brings together novel and more traditional methods in molecular biology and immunology, in order to provide a tool-kit for all stages for Celiac Disease research-from the practicalities of identifying, obtaining, stabilizing and maintaining high quality samples, to molecular analysis and bioinformatics.

This detailed volume provides an overview of recent techniques employed in the field of G protein-coupled receptors (GPCRs) to screen for new drugs and to derive information about their receptor structure, dynamics, and function for the purpose of developing improved therapeutics. Owing to remarkable recent advances in the structural, biophysical and biochemical analyses of these receptors, as well as a growing body of evidence hinting at the possible relevance of allosteric modulators, biased agonists and oligomer-selective ligands as improved therapeutic agents, drug discovery for GPCRs has recently taken a completely new direction. For this book, expert contributors have shared their protocols and views on the impact of these methodologies on modern drug discovery. 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.Practical and fully updated, G Protein-Coupled Receptors in Drug Discovery: Methods and Protocols, Second Edition serves as an ideal guide for a diverse audience from structural and molecular biologists to pharmacologists and drug designers who wish to explore this extensive class of key drug targets.

This volume expands upon Protein Electrophoresis (2012) and provides readers with easy-to-follow and reproducible methods to study electrophoresis. The chapters in this book cover topics such as the Cydex Blue assay; cellulose-acetate electrophoresis of hemoglobin; cationic electrophoresis; tricine-SDS-Page; identification of proteins on archived 2-D gels; cell surface protein biotinylation of SDS-PAGE analysis; and artifacts and common errors in protein gel 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.Practical and thorough, Electrophoretic Separation of Proteins: Methods and Protocols is a valuable resource for researchers who are interested in learning and experimenting with this field.