Böcker utgivna av Springer-Verlag New York Inc.

The aim of this volume is to provide a comprehensive description of methods and protocols useful for the further study of T-helper cells. Chapters guide readers through T-helper cell recovery, molecular study, signal transduction pathways, T-cell manipulation and, last but not least, ¿omic¿ approaches. 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, T- Helper Cells: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field.

This detailed book aims to provide an overview of novel bioinformatics resources for exploring diverse aspects of RNA biology. This edition focuses on methods dealing with non-coding RNA (miRNAs, circRNAs or lncRNAs), RNA modifications (m6A or RNA editing), single cell RNA-seq and statistical models to handle count data from RNA-seq experiments. The book also includes chapters based on the classical RNA bioinformatics methods, such as those for deciphering secondary and tertiary RNA structures; however, they are revised to take into account deep sequencing data. Finally, chapters describing methods to analyze RNA sequencing data from emerging third generation sequencing technologies that could provide interesting insights into the transcriptional process and its regulation are also included. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of expert implementation advice that encourages quality results. Comprehensive and up-to-date, RNA Bioinformatics, Second Edition serves as an ideal guide for researchers digging ever-deeper into the depths of the study of RNAs.The chapter 'RNA-Seq Data Analysis in Galaxy' is open access under a CC BY 4.0 license.

This volume details protocols on rationale design of therapeutic siRNA molecules and its encapsulation with smart vehicles to overcome the barriers to an effective administration in vivo. 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, Design and Delivery of SiRNA Therapeutics aims to ensure successful results in the further study of this vital field.

This volume explores the latest methods used to study various aspects of TET proteins and their biology. Chapters in this book are divided into five parts. Part One describes technologies aimed at detecting and quantifying DNA methylation turnover using massively parallel sequencing, ELISA, and mass spectrometry approaches. Part Two looks at data analyses protocols for distinguishing acting versus passive DNA demethylation and estimation of 5mC and 5hmC levels. Part Three deals with a new topic that takes advantage of modified CRISPR/Cas9 genome editing systems to target DNA demethylation activity to genomic loci of interest. Part Four discusses protocols that detail how to purify TET proteins and unravel their protein interactions, and Part Five looks at the assessment of TET protein function and activity in vivo and in vitro. 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, TET Proteins and DNA Demethylation: Methods and Protocols is a valuable resource that aims to help research scientists at all levels working in the fields of DNA demethylation dynamics.Chapters 3, 7 and 17 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume provides a comprehensive set of protocols that can be used by any research lab to investigate diverse functional and structural properties of Single Stranded DNA Binding Proteins (SSBs) from eubacterial, archaeal, eukaryotic, mitochondrial and viral 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, Single Stranded DNA Binding Proteins aims to be a useful practical guide to researchers to help further their study in this field.

This detailed book highlights recent advances in molecular imaging techniques and protocols, designed to be immediately applicable in global bio-laboratories. The chapters are categorized into seven major groups according to the reporter materials, such as imaging with passive optical readouts, activatable bioluminescent probes, functional substrates and luciferases, organic fluorescent probes, BRET probes, FRET probes, as well as with advanced instrumentation. 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, Live Cell Imaging: Methods and Protocols aims to direct and inspire researchers into creating smarter, next-generation imaging techniques that are truly quantitative, highly sensitive, and readily comprehended, in the effort to engender deeper understanding of biological systems and break new ground in the research fields of life science.

This thorough book explores some of the most important methods and concepts affecting the quantitative analysis of the transport, targeting, and disposition of chemicals within cells, which in turn impact the macroscopic pharmacokinetics of chemical agents in the whole organism. The first half of the volume focuses on small organic molecules with drug-like characteristics, while the second half delves into the cellular pharmacokinetics of biologics and other macromolecules, including peptide therapeutics, cyclotides, antibodies, as well as nanoparticles, thus creating a comprehensive treatise that approaches cellular pharmacokinetics from the different perspectives of pharmaceutical scientists, chemical biologists, medicinal chemists, and protein engineers dealing with very different chemical agents spanning a wide range of sizes, physicochemical properties, and targeting mechanisms. Written for the Methods in Pharmacology and Toxicology series, chapters provide the kind of key detail and expert implementation advice that leads to excellent results in the lab. Synthetic biologists, biophysicists, and bioengineers are amongst the long list of scientists who could benefit from reading this book or from using it as a textbook. Authoritative and practical, Quantitative Analysis of Cellular Drug Transport, Disposition, and Delivery builds on a long history of drug development and the adding of quantitative methods at the cellular scale in order to inspire new approaches to drug development that are better able to take advantage of phenomena such as soluble-to-insoluble phase transitions or bispecific targeting, which could ultimately be exploited for the development of more effective drug delivery systems and therapeutic agents.

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.