Books in the Methods in Molecular Biology series

This volume explores lipidomics through protocols that focus on areas of utility, techniques, and bioinformatics advancements. The protocols in this book cover topics such as isolation of specific membranes and specialized fractionation of subcellular compartments, and computational and functional analysis of lipid metabolizing enzymes. Each protocol also discusses methods of dealing with fractionated organelles¿cells to whole organisms. 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, Lipidomics: Methods and Protocols is a useful and valuable resource for biochemists, molecular biologists, and neuroscientists interested in neurology, ophthalmology and vision sciences, as well as mass spectrometrists with interest in disease discovery.

This volume presents established bioinformatics tools and databases for function prediction of proteins. Reflecting the diversity of this active field in bioinformatics, the chapters in this book discuss a variety of tools and resources such as sequence-, structure-, systems-, and interaction-based function prediction methods, tools for functional analysis of metagenomics data, detecting moonlighting-proteins, sub-cellular localization prediction, and pathway and comparative genomics databases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, step-by-step instructions of how to use software and web resources, use cases, and tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Protein Function Prediction: Methods and Protocols is a valuable and practical guide for using bioinformatics tools for investigating protein function

This volume discusses protocols that aid in measuring different components in the renin-angiotensin-aldosterone system (RAAS). The book also looks at the methods used to assess angiotensin peptides and discerning the influence of RAAS components on different mammalian diseases. The chapters cover topics such as the use of fluorescent substrate to measure ACE2 activity in the mouse abdominal aorta, blood pressure monitoring using the radio telemetry method, and the analysis of angiotensin metabolism in the kidney using mass spectrometry. 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, The Renin-Angiotensin-Aldosterone Systems: Methods and Protocols is a valuable resource that provides scientists and researchers with the best approaches to examine RAAS.

This volume details protocols that cover the broad arsenal of techniques used to study a secretion system from A to Z. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as reporters to follow effector transport. 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, Bacterial Protein Secretion Systems: Methods and Protocol aims to provide techniques that are not restricted to the study of secretion systems but are also of specific interest for any researcher interested on multi-protein complexes of the bacterial cell envelope.

This volume addresses the various techniques and novel applications of mass spectrometry imaging (MSI) and its role as a discovery tool in the field of proteomics, lipidomics, and metabolomics. The chapters in this book demonstrate how MSI can be applied to many areas of research such as clinical pathology, translational medicine, toxicology, biomarkers and response studies, and potential incorporation of MSI into forensic workflows. 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. Innovative and comprehensive, Imaging Mass Spectrometry: Methods and Protocols is a valuable resource for research scientists and clinicians who are interested in further studies of MSI technologies.

This volume details PCR technique with focus on its application specificities to the biotechnology and bioengineering field. Chapters are broken into five sections covering sgeneral PCR protocols, different applied examples to molecular and synthetic biotechnology, food science and technology, environmental microbiology and molecular ecology, and healthcare. 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, PCR: Methods and Protocols hopes to be a valuable and useful resource for wet lab researchers, particularly within the biotechnology and bioengineering field.

Chapters guide readers through comprehensive experimental approach for molecular investigations of plant receptor kinases, from the simplest methods for expression and purification of receptor domains to the most advanced methods aiming at understanding the dynamics of receptor complex formation and specificity of signaling pathways.

This vital book collects key methods that have supported advancements in the field of ebolavirus molecular biology given the pressing need for the advancement of techniques for diagnostics, the development of vaccines and antivirals, and for furthering our understanding of ebolavirus biology.

This volume presents protocols that analyze and explore gene regulatory networks (GRNs) at different levels in plants. This book is divided into two parts: Part I introduces different experimental techniques used to study genes and their regulatory interactions in plants. Part II highlights different computational approaches used for the integration of experimental data and bioinformatics-based predictions of regulatory interactions. This part of the book also provides information on essential database resources that grant access to gene-regulatory and molecular interactions in different plant genomes, with a specific focus on Arabidopsis thaliana. 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, Plant Gene Regulatory Networks: Methods and Protocols is a valuable resource for scientists and researchers interested in expanding their knowledge of GRNs.

This volume details protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. 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 practical, Genome Editing in Animals: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This volume describes up-to-date techniques used in understanding the molecular biology of acute myeloid leukemia (AML) and addressing advances in diagnosis, classification, prognostication, and therapeutic strategies to potentially impact overall patient survival.

Thisvolume provides a variety of protocols to analyze various epigeneticchanges, including differential expression of non-coding RNAs, changes in DNAmethylation, and histone modifications in plants. Chapters detail protocolswith different degrees of complexity, and describe bioinformatics approachesfor data processing and analysis. 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 andcutting-edge, Plant Epigenetics: Methods and Protocols, Second Editionaims to ensure successful results in the further study of this vital field.

This volume aims to provide a range of methods and protocols for studying tumor angiogenesis in vitro and in vivo to reflect advances in the field. The chapters in this book cover topics such as: morphological aspects of tumor angiogenesis, aortic ring assay and its use for the study of tumor angiogenesis, ex vivo tissue culture model for anti-angiogenic drug testing, transgenic zebrafish, orthotopic models of ovarian cancer, and uncovering metabolic effects of anti-angiogenic therapy in tumors by induced metabolic bioluminescence imaging. 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, Tumor Angiogenesis Assays: Methods and Protocols is a valuable resource for anyone interested in tumor angiogenesis assay research.

This volume presents state-of-the art methods for the synthesis, design, assembly, post synthesis processing, and application of synthetic DNA to modern biotechnology.

This volume presents the most recent technologies used in the Polycomb Group Proteins (PcG) field. Chapters detail state-of-the-art methods, creating a unique and comprehensive reference source for investigating Polycomb function in the nucleus. 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, Polycomb Group Proteins: Method and Protocols aims to ensure successful results in the further study of this vital field.

This second edition expands on the previous edition with new chapters that are suitable for newcomers, as well as more detailed chapters that cover protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the challenges that scale-up of production poses to the investigator. Many of the chapters also discuss generation and purification of recombinant proteins, recombinant antibody production, and the tagging of proteins as a means to enhance their solubility and simplify their purification on an individual scale or in high-throughput systems. This book also provides readers with chapters that describe not just the more commonly used methods, but also recently developed approaches such as proteomic/mass spectrometric techniques and Lectin-based affinity chromatography. 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, Protein Chromatography: Methods and Protocols, Second Edition is a valuable resource for anyone who is interested in the field of protein chromatography.

This volume provides a collection of techniques and approaches for the study of ERK signaling. It begins with a historical perspective of genetic and molecular discoveries, followed by chapters covering specific topics in a broad range of experimental systems, including in vitro assays of EGFR and ERK activities; proteomic and genome-wide analyses of ERK signaling targets; cell biological, genetic, quantitative and imaging approaches in cells and model organisms; and integrated mathematical modeling of the ERK pathway. 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, ERK Signaling: Methods and Protocols will aid and stimulate further advances in the vibrant field of ERK signaling.

This volume focuses on the use of system genetic methods and the use of murine models to study the role of gene variants and environmental factors on human health and disease-what is now often called personalized or precision health care.

This volume provides a comprehensive guide on the Cyr61 (Cysteine-rich 61) (CCN) family of proteins and genes from basic research to cutting-edge methodologies and state-of-the-art techniques.

This volume provides protocols and procedures for determining and modeling RNA structure. Chapters guide the reader through protocols for RNA secondary structure prediction, single sequence modeling, Crumple, RNAstructure to model conserved secondary structures with multiple homologs, the prediction of bimolecular secondary structures with RNAstructure, STarMir, protocols for structure mapping, mapping data to constrain or restrain RNA secondary structure prediction with RNAstructure, unassigned NMR resonances, modeling protocols for Rosetta FARFAR, RNAComposer , ModeRNA, and MC-Fold. 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 Practical, RNA Structure Determination: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This volume focuses on applications of split inteins, and the progress that has been made in the past 5 years on discovery and engineering of fast and more efficient split inteins. The first few chapters in Split Inteins: Methods and Protocols explore new techniques on how to use split inteins for affinity purification of overproduced proteins, and split-intein based technologies to prepare cyclic peptides and proteins. The next few chapters discuss semisynthetic protein trans-splicing using one synthetic intein piece, synthetic intein-extein pieces used to deliver other cargos for chemical modification both of purified proteins and of proteins in living cells, as well as isotopic labeling of proteins for NMR studies, and a discussion on how protein block copolymers can be generated by protein trans-splicing to form protein hydrogels. The last few chapters deal with intein applications in transgenic plants and conditional inteins that can be regulated in artificial ways by small molecules or light, a cassette-based approach to quickly test many intein insertion positions, and a computational approach to predict new intein split sites (the approach also works for other proteins). Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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, Split Inteins: Methods and Protocols is a valuable resource that will provide guidance toward possibilities of split intein applications, explore proven and detailed protocols adaptable to various research projects, and inspire new method developments.

This detailed volume presents a comprehensive technical overview of DNA nanotechnology with an emphasis on 3D DNA nanostructure design and applications. Coverage spans from basic design principles for DNA and RNA nanostructures to their cutting-edge applications in a variety of fields, with the book divided into basic DNA and RNA nanostructure design strategies as well as applications utilizing DNA nanostructures, including but not limited to nanomedicine, bioimaging, biosensing, nanoplasmonics, nanoelectronics, nanofabrication, crystallography, biophysics, and analytical chemistry. 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. Comprehensive and authoritative, 3D DNA Nanostructure: Methods and Protocols provides the most up-to-date tutorial style overviews and technical style protocols to benefit researchers in a wide variety of areas.

In Vitro Mutagenesis: Methods and Protocols guides readers through methods for gene and genome editing, practical bioinformatics approaches for identifying mutagenesis targets, and novel site-directed and random mutagenesis approaches aimed at gaining a better understanding of protein-protein and protein-cofactor interactions.

This volume provides an overview of the field and practical hints for vaccinologists in academia and industry. Chapters provide protocols on self-replicating RNA vectors, non-replication mRNA vectors, adjuvantation and delivery, and preclinical and clinical development. 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, RNA Vaccines: Methods and Protocols aims to increased collaboration on RNA vaccines between basic and applied scientists in academia, government, and industry to develop future solutions for today¿s challenges.

This detailed book collects the main methodologies used for the analysis of the activity, localization, and regulation of the components of the Mitotic Exit Network (MEN) pathway during mitotic exit in Saccharomyces cerevisiae, as well as for the evaluation of the roles of these proteins in other cellular processes, such as the condensation of the rDNA, the functionality of the mitotic checkpoints, and cytokinesis. Budding yeast serves as an ideal model system for dissecting the mechanisms that regulate cell cycle progression and providing new insights into the molecular basis of cell cycle control and, thus, into the origin of diseases that arise as a consequence of problems during cell division. Therefore, although this volume concentrates on Saccharomyces cerevisiae as a model, it also details the implications that the research about the MEN have on our understanding of the mitotic exit process in higher eukaryotes. 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, The Mitotic Exit Network: Methods and Protocols will be a valuable reference for cellular and molecular biologists and biochemists as well as for all scientists interested in the study of the regulation of mitotic exit using budding yeast as a model organism.

This volume features up-to-date protocols for the isolation, preservation, and validation of various cell sources comprising large and small animal models, examining the impact of cell transplantation on acute and chronic liver diseases.

This detailed volume presents timely and authoritative content offering a comprehensive overview of the current state of the art in fungal diagnostics. Moreover, it addresses on-going developments expected to provide a basis for targeted treatment strategies resulting in improved outcome of invasive mycoses. The knowledge of host-related predisposing factors and stratified treatment options facilitating timely onset of adequate antifungal therapy are critical for successful clinical management and outcome of invasive fungal disease (IFD), requiring not only rapid diagnosis of a fungal infection and identification of the causative species, but also assessment of pathogen/host factors related to pathogenicity, susceptibility, and response to treatment. 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 protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, Human Fungal Pathogen Identification: Methods and Protocols serves as an ideal reference for researchers investigating the ever-growing worldwide healthcare problems involving fungal infections.

This book brings together the major techniques used in the isolation or enrichment of individual populations of organelles and other subcellular structures from plants with the goal that, by being able to isolate subcellular structures, the research and understanding of various facets of compartmentalized function in plant cells can be advanced.

This volume is a valuable and timely resource for a broad audience with interests in basic and translational cancer biology, cancer drug development, as well as in the practice of personalized oncology.

The volume provides comprehensive, state-of-the-art experimental techniques that are now available to dissect the molecular mechanisms of regulation and function of cohesin and the related factor condensin in vitro and in vivo across different model organisms, as well as in human cells.