Bücher veröffentlicht von Springer-Verlag New York Inc.

This detailed book collects original protocols aimed at encouraging and stimulating the scientific community to design and produce models for the laboratory that mimic cell guidance conditions as they occur in vivo. The protocols collected describe powerful strategies to exploit chemical cues involved in cell differentiation processes. Special emphasis is given to the use of methods for purification and characterization of exosomes and other secreted vesicles, as well as micro and non-coding RNAs, that have been demonstrated to control the tuning of the in vivo micro and macro environment in order to ensure the optimal soluble environment in vitro. 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, Next Generation Culture Platforms for Reliable In Vitro Models: Methods and Protocols serves as an ideal guide for researchers working toward developing these vital testing models for preclinical studies.

This volume presents methods used for the analysis of glycoproteins at different levels¿intact, subunit, glycopeptide, and monosaccharide--, and discusses and solves most analytical challenges that a scientist working on glycoproteins may come across. The chapters in this book cover topics such as the role of glycosylation on the properties of therapeutic glycoproteins; different analytical methods to characterize glycosylation, from the intact proteins to the glycan level, for both N-linked and O-linked glycoproteins; mass spectrometry imaging methodology for glycosylation analysis in tissues; approaches to characterizing glycosylation on cultured cells; and the use of cloud computing to deploy mass spectrometry data analysis. 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, Mass Spectrometry of Glycoproteins: Methods and Protocols is a valuable resource for scientists interested in learning more about this developing field.

This second edition provides new and updated chapters useful for the study of Regulatory B cells. Organized in four sections, chapters detail basic methods for the isolation and immunophenotypical analysis of these cells, experimental approaches for the ex vivo generation/expansion of IL-10 producing B cells, molecular biology techniques for the analysis of IL-10 expression and production, and animal models mimicking pathologic settings. Written in the highly successful Methods in Molecular Biology series format, chapters include an introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Regulatory B Cells: Methods and Protocols, Second Edition aims to be useful to the scientific community and serve toclarify some unsolved aspects of Regulatory B Cells research.

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.