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This volume explores the latest advancements in the field of cell cycle checkpoints and their implications for human diseases. Chapters in this book cover topics such as post-translationally modified p53 by western blotting; CHK1 cellular localization by immunofluorescence microscopy; DNA affinity purification; knockdown of target genes by siRNA in vitro; and calreticulin exposure in mitotic catastrophe. 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, Cell Cycle Checkpoints: Methods and Protocols is a valuable resource for researcher interested in learning more about this developing field.
This volume explores the recent advances in the study of translational paths in central inflammation and focuses on ongoing pathophysiological processes and the transition between inflammatory stages and progressive states with neurodegeneration. Chapters cover topics such as pathophysiological hallmarks of neuroinflammation from tissue damage to reorganization; connecting studies of mouse models; and investigations of humans with multiple sclerosis. In the 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 practical, Translational Methods for Multiple Sclerosis Research is a valuable resource for researchers who want to learn more about this chronic ad progressive disease, and pave the way for new advancements.
This third edition provides new and updated chapters detailing a complete introduction to common and emerging procedures for characterizing the interactions of individual proteins with their natural ligands, drugs or other binding partners. Chapters detail natural substrates, potential drug leads, quantitative understanding of the mechanism of interaction, and different 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. Authoritative and accessible, Protein Ligand Interactions: Methods and Applications, Third Edition serves as an ideal guide for researchers new to the field of biophysical characterization of protein interactions.
This volume compiles new experimental approaches and concepts focusing mostly, but not solely, on ways to manipulate and regulate Ras activity and its downstream signaling output. Chapters detail standard methodologies, biochemical methods, Ras processing trafficking and localization, Ras signaling and inhibition, and in vivo models for studying Ras function. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Ras Activity and Signaling: Methods and Protocols aims to provide support and guidance to lab workers in their work on Ras GTPases and in the design of new projects requiring novel methodologies.
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 detailed volume presents a comprehensive bioinformatic and experimental toolbox for prioritizing, annotating, and functionally analyzing long non-coding RNAs (lncRNAs). Playing a vital role in diverse biological progresses and human disease, lncRNAs have proven to be a challenging subject of study due to our limited understanding of their sequence-function relationships, lack of complete genetic annotation, and the unavailability of systems required to define their functional importance and molecular mechanisms, all of which this book seeks to address. 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, Functional Analysis of Long Non-Coding RNAs: Methods and Protocols provides atimely and convenient resource to facilitate the identification and characterization of disease-associated human lncRNAs, which aims to shed light on their role in biology and pathophysiology and ultimately lead toward novel therapeutic approaches targeting lncRNAs for the amelioration of human diseases.
This book provides a single-source reference on the current state of the ribosome profiling method by describing experimental protocols for the quantitative analysis of translation in a variety of model organisms. In addition, the volume presents a detailed overview of the existing software tools and includes detailed description of methods for statistical analysis, data processing, and visualization of ribosome profiling data. 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, Ribosome Profiling: Methods and Protocols aims to provide the type of standardized protocols that have previously been unavailable in an effort to bypass the major barriers to wide use of ribosome profiling-based approaches.
This detailed book examines experimental approaches used to investigate the regulation and function of phosphoinositides (PtdInsP), rare eukaryotic phospholipids with a broad role in biological processes such as signal transduction, cell migration and adhesion, cell growth, subcellular organization, and membrane trafficking. The combination of complementary biochemical, mass spectrometry, and imaging methods are instrumental for the detection and quantification of PtdInsP species, as well as induced dimerization methods, affinity precipitation or co-sedimentation with liposomes, protein insertion within lipid bilayers, and enzymatic assays, or through emerging methods like native mass spectrometry and microfluidics, all of which are touched upon in this volume. 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 laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Phosphoinositides: Methods and Protocols provides detailed methodology for both specialist and novice researchers on a variety of complementary methods that have been instrumental in dissecting the regulation, dynamics, and function of PtdInsPs.
This third edition volume expands on the previous editions with updated chapters on longitudinal studies, randomized trials, evidence-based decisions making, and a new section on changing health-related behaviors. The chapters in this book are organized into six parts: Part One focuses on framing clinical research questions and choosing a suitable design; biases that may occur in clinical research; and the ethics associated with doing conducting research on humans. Parts Two through Four discuss designs, measurements, and analysis that pertain to evaluation of risk in longitudinal studies; assessment of therapy in controlled trials; and evaluation of diagnostic tests. Part Five presents methods used in various components of evidence-based decision-making; and Part Six highlights interventions focused on changing health-related behaviors. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of various types of bias, step-by-step, readily reproducible protocols for different research designs, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, Clinical Epidemiology: Methods and Protocols, Third Edition is a valuable resource for clinicians and researchers who want to expand their works to humans and use their findings in the health system.
This volume details the most up-to-date technologies used in plant transposable element studies and provides easy-to-follow protocols. Chapters guide readers on available database resources, annotation of different families of transposon, and experimental methods to detect their transposition intermediates, neo-transposed DNAs, and transposition events. 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 Transposable Elements: Methods and Protocols aims to provide web-lab and dry-lab methodologies targeted at various levels from beginner to experienced.
This volume explores strategies and detailed protocols for the preparation of macromolecular complexes and their characterization in view of structural analysis. The chapters in this book are separated into three parts: Part One focuses on sample preparation, and covers strategies for recombinant expression of multiprotein complexes in prokaryotic and eukaryotic hosts, for genome engineering using the CRISPR/Cas9 system and for production of specific binders such as reformatted antibodies and artificial binding proteins. Part Two looks at the biophysical methods that can provide useful indicators for sample optimization, and often complement structural information obtained with core technologies for structure determination¿x-ray crystallography and cryo-electron microscopy¿by quantitative solution data. Part Three discusses the characterization of multiprotein complexes in a cellular environment using the latest technologies and in vivo approaches. Written in the highly successfulMethods 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, Multiprotein Complexes: Methods and Protocols is a valuable resource for structural and molecular biologists who need to prepare multi-components for their applications, and for other scientists working on macromolecular assemblies from other angles that need to know the latest approaches that the field has to offer.
This volume explores strategies and detailed protocols for the preparation of macromolecular complexes and their characterization in view of structural analysis. The chapters in this book are separated into three parts: Part One focuses on sample preparation, and covers strategies for recombinant expression of multiprotein complexes in prokaryotic and eukaryotic hosts, for genome engineering using the CRISPR/Cas9 system and for production of specific binders such as reformatted antibodies and artificial binding proteins. Part Two looks at the biophysical methods that can provide useful indicators for sample optimization, and often complement structural information obtained with core technologies for structure determination¿x-ray crystallography and cryo-electron microscopy¿by quantitative solution data. Part Three discusses the characterization of multiprotein complexes in a cellular environment using the latest technologies and in vivo approaches. Written in the highly successfulMethods 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, Multiprotein Complexes: Methods and Protocols is a valuable resource for structural and molecular biologists who need to prepare multi-components for their applications, and for other scientists working on macromolecular assemblies from other angles that need to know the latest approaches that the field has to offer.
This detailed book collects a variety of methods from investigators at the forefront of chondrocyte biology and pathology research that they have tailored for their research projects in order to tackle the unique properties and challenges of the chondrocyte and those of its tissue matrix. Divided into three parts, the volume explores experimental models to study chondrocytes, in vivo assays, as well as methods to characterize the chondrocyte phenotype with high-throughput or discrete assays on cells maintained in vivo, just isolated from their tissue (ex vivo), or cultured in vitro. Written in 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 cutting-edge, Chondrocytes: Methods andProtocols is an ideal guide for experts as well as newcomers in the cartilage field with the aim of sparking new research ideas in order to solve many of the pending mysteries regarding cartilage biology, disease mechanisms, and potential treatments.
This volume provides a comprehensive review of concepts and protocols related to fluorescence in situ hybridization (FISH) applied to microbial cells. Chapters will serve guide for the design of new probes and the development of novel FISH-based protocols. 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, Fluorescence In Situ Hybridization (FISH) for Microbial Cells: Methods and Concepts aims to ensure successful results in the further study of this vital field.
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