Due to the vital biological importance of RNA and proteins functioning together within a cell, a protocol volume describing experimental procedures to study their interactions should find a home in many laboratories. RNA-Protein Interaction Protocols, Second Edition updates, complements, and expands upon the popular first edition by providing a collection of cutting-edge techniques developed or refined in the past few years along with tried-and-true methods. The expert contributors explore the isolation and characterization of RNA-protein complexes, the analysis and measurement of RNA-protein interaction, and related novel techniques and strategies. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, RNA-Protein Interaction Protocols, Second Edition is an ideal guide for researchers continuing the study of this all-important biological partnership.

This book is specifically about the application of the extremely powerful interaction between the protein avidin or its homologues and the vitamin biotin and some of its homologues. With excellent descriptions of laboratory protocols written by expert researchers, this volume is equally perfect for the student or the professional laboratory scientist.

In this updated volume, experts from around the world provide the latest protocols for isolating different organelles and the localization of particular proteins using a variety of methods such as light, confocal, and electron microscopy. Emphasis is placed on protein targeting of cellular compartments in both prokaryotic and eukaryotic systems. The book includes targeting protocols from different systems.

This is a fully up-dated and expanded practical guide to protein structure-function relationships. This important area of research is brought up-to-date by the leading scientists in the field. The compilation of detailed protocols focuses on protein function, proteome research and characterization of pharmaceutical proteins, while following the successful format of the Methods in Molecular Biology series. Comprehensive and cutting edge, the book serves as practical guide for researchers working in the field of protein structure-function relationships and the rapidly growing field of proteomics, as well as scientists in the pharmaceutical industries.

This book covers elements of both the data-driven comparative modeling approach to structure prediction and also recent attempts to simulate folding using explicit or simplified models. Despite the unsolved mystery of how a protein folds, advances are being made in predicting the interactions of proteins with other molecules. Also rapidly advancing are the methods for solving the inverse folding problem, the problem of finding a sequence to fit a structure.

This book focuses on the various computational methods for prediction, their successes and their limitations, from the perspective of their most well known practitioners.

This is the first book to examine organelle proteomics in depth.

It begins by introducing the different analytical strategies developed and successfully utilized to study organelle proteomes, and detailing the use of multidimensional liquid chromatography coupled to tandem mass spectrometry for peptide sample analysis. Detailed protocols are provided and a section is devoted to methods enabling a global estimate of the reliability of the protein list assigned to an organelle.

In one volume this book provides useful and innovative protocols developed specifically for the proteomic profiling of human tissues. The book provides high-throughput gel-based techniques, microarrays and a number of other methods used in proteomic research. This important book will prove indispensable to investigators of biomarker discovery and therapeutic response profiling, as well as those forging new paths in the fields of theranostics and personalized medicine.

As research has progressed, the cannabinoid CB 1 and CB 2 receptors have expanded significantly in importance within the neuroscience mainstream. In The Cannabinoid Receptors, leading experts introduce newcomers to the cannabinoid field with chapters covering cannabinoid ligand synthesis and structure activity relationships, the molecular pharmacology of the cannabinoid receptors and the endocannabinoid system, and ultimately, the whole animal pharmacology and therapeutic applications for cannabinoid drugs. Adding to those key topics, the book also examines the current direction of the field with chapters on new putative cannabinoid receptors and challenges for future research. As a part of The Receptors series, this volume highlights its receptor with the most thorough, focused and essential information available.

Comprehensive and cutting-edge, The Cannabinoid Receptors serves as an ideal guidebook to what continues to be a fascinating and vital field.

Post-translational protein modifications by members of the ubiquitin family are widely recognized as important regulatory control systems for a variety of biological pathways. Their influence on eukaryotic cellular metabolism is comparable to that of other modifi- tions such as phosphorylation, acetylation and methylation. The small ubiquitin-related modifier SUMO uses a conjugation and de-conjugation system closely related to that of ubiquitin itself; yet, the functions of the SUMO system are highly diverse and largely independent of the ubiquitin system. SUMO modification controls the activity of tr- scription factors and can influence protein stability, but it also contributes to nucleo-cy- plasmic transport, chromosome segregation and DNA damage repair. As a consequence, the SUMO system pervades virtually all areas of basic molecular and cell biology, and scientists from different backgrounds, including medical researchers, are likely to enco- ter SUMO in the course of their studies. This volume, SUMO Protocols, therefore aims at presenting a collection of methods relevant to SUMO research in order to make these tools available to biochemists, molecular and cell biologists as well as research-oriented clinicians not yet familiar with the system. In contrast to the ubiquitin system, which has been the subject of several reviews and methods collections, no practical compendium entirely devoted to SUMO has been p- lished.

Gene expression can mean the difference between a functional and non-functional genome, between health and disease, and with the development of transgenic crops, the difference between survival and starvation. In DNA-Protein Interactions: Principles and Protocols, Third Edition, this vital subject is brought up to date with protocols exploring the most cutting-edge developments in the field, including in vivo and genome-wide interaction techniques. Addressing topics such as chromatin immunoprecipitation, topological studies, photocrosslinking, FRET and imaging techniques, the volume fully updates and expands upon the successful previous editions. Written in the convenient and informative Methods in Molecular Biology (TM) series format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, DNA-Protein Interactions: Principles and Protocols, Third Edition serves as an ideal guide for all those exploring this dynamic, essential, and increasingly affordable area of research.

Over the course of the past decade, there have been remarkable advances in the study of human pathogenic fungi. These developments have taken place throughout a wide range of disciplines, and have come as the result of newly available genome sequences of pathogens such as candida albicans and other model fungi. In Candida Albicans: Methods and Protocols, expert researchers explore these exciting new insights, focusing on the study of medically important fungi and Candida spp in particular. Chapters examine critical aspects of molecular methods, providing information on reporter gene assays, transformation, gene expression in vivo, and methods for large-scale gene disruption. At the same time, the work includes in-depth descriptions of disease models of candidiasis, facts about strain identification, and guidelines on the preparation of samples for proteomic investigations and tandem affinity purification. Composed in the highly successful "Methods in Molecular Biology " series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting edge, Candida Albicans: Methods and Protocols is an invaluable source of methods for investigators in the exhilarating fields of medical and molecular mycology."

This book is indexed in Chemical Abstracts ServiceThe interactions of proteins with other molecules are important in many cellular activities. Investigations have been carried out to understand the recognition mechanism, identify the binding sites, analyze the the binding affinity of complexes, and study the influence of mutations on diseases. Protein interactions are also crucial in structure-based drug design.This book covers computational analysis of protein-protein, protein-nucleic acid and protein-ligand interactions and their applications. It provides up-to-date information and the latest developments from experts in the field, using illustrations to explain the key concepts and applications. This volume can serve as a single source on comparative studies of proteins interacting with proteins/DNAs/RNAs/carbohydrates and small molecules.

The book helps the reader to better understand cytogenetics and the intricacies of the methodology. The different methods of fluorescence in situ hybridization are discussed and the results achieved are presented. The book provides a comprehensive review of basic and applied aspects of cytogenetics and thus is of intense interest to all those interested in chromosomes and their alterations by different types of mutagens, including chemical mutagens and ionizing and nonionizing radiation, with special reference to electromagnetic fields.

Researchers in structural genomics continue to search for biochemical and cellular functions of proteins as well as the ways in which proteins assemble into functional pathways and networks using either experimental or computational approaches. Based on the experience of leading international experts, Structural Genomics and High Throughput Structural Biology details state-of-the-art analytical and computational methods used to reveal the three-dimensional structure and function of proteins. A historical perspective and a detailed guide to the production of protein material for structural determination, a key step in the process, lay the necessary foundation for discussing the most effective structure determination technologies, such as X-ray crystallography and NMR spectroscopy. Encouraging the study of genes and proteins of unknown structure in order to discover new information about folding, specific structural features, or function, Structural Genomics and High Throughput Structural Biology presents the methods used to interpret the sequences of proteins in a structural context, giving insight into their function. It also explains how to extract information from public data repositories and how to account for variability and accuracy in the quality of this data. The book concludes with a discussion of practical applications of therapeutically driven structural genomics, and presents future directions in the field. Structural Genomics and High Throughput Structural Biology offers a comprehensive guide to the theoretical, technological, and experimental methodologies used to derive structural information from encoded proteins by renowned and world leading scientists in the field.

First methods book which includes many detailed descriptions

Absolutely needed and thus timely for the scientific community

Comprises 15% more content and includes the mentioned special features

First introduced to biomedical research in 1980, the term biomarker has taken on a life of its own in recent years and has come to mean a number of things. In biomedical science, biomarker has evolved to most commonly mean a characteristic that can be used either as a diagnostic or a prognostic, but most significantly as a screening indicator for pathologies that tend to be somewhat silent prior to overt clinical display. Applying scientific rigor, as well as a disciplined approach to nomenclature, Roger Lundblad's Development and Application of Biomarkers rationalizes the current enthusiasm for biomarkers with the use of well-established clinical laboratory analytes in clinical medicine. Highly respected for his work as both a classical protein scientist and as a pioneer in proteomics, Dr. Lundblad catalogs various biomarkers recognized in clinical medicine and, where possible, matches the expectations for advances in screening technologies with the realities of statistical analysis. More specifically, this important reference: Details an extensive list of biomarkers for various stages of a number of cancer types including ovarian, pancreatic, prostate, and breast cancer Looks at how proteomics is used for the discovery and validation of biomarkers Explores the use of microarray technology, ultra-high performance liquid chromatography, and computational bioinformatic approaches for the discovery and use of biomarkers Examines the use of cells and cell fragments as more complex biomarkers Organizes a host of significant biomarkers and essential research by type and use in a series of readily accessible tables Throughout this volume, Dr. Lundblad encourages consideration of biomarkers more as a concept than as laboratory analytes, emphasizing the relation between the discovery of a biomarker and the biology underlying its production. Ultimately, it is a thorough understanding of that underlying biology that will lead to the development of assays that are robust and reproducible, as well as clinically significant.

The Handbook of Biomedical Nonlinear Optical Microscopies provides comprehensive treatment of the theories, techniques, and biomedical applications of nonlinear optics and microscopy for cell biologists, life scientists, biomedical engineers, and clinicians. The chapters are separated into basic and advanced sections, and provide both textual and graphical illustrations of all key concepts. The more basic sections are aimed at life scientists without advanced training in physics and mathematics, and tutorials are provided for the more challenging sections. The first part of the Handbook introduces the historical context of nonlinear microscopy. The second part presents the nonlinear optical theory of two- and multiphoton excited fluorescence (TPE, MPE) spectroscopy, second and third harmonic generation (SHG, THG) spectroscopy, and coherent anti-Stokes Raman spectroscopy (CARS). The third part introduces modern microscopic and spectroscopic instrumentation and techniques that are based on nonlinear optics. The fourth part provides key applications of nonlinear microscopy to the biomedical area: neurobiology, immunology, tumor biology, developmental biology, dermatology, and cellular metabolism. There are also chapters on nonlinear molecular probes, cellular damage, and nanoprocessing.

How does the chemical description of a protein relate to its three-dimensional structure? How does the three-dimensional structure relate to the machinery that brings about a chemical reaction? How does the sequence of a gene encode not only the sequence of a protein but, more importantly, the architecture and function of that protein? Protein Structure and Function provides a clear and critical survey of our current understanding of the structure and function of proteins to answer questions such as these. Opening with a consideration of the link between protein sequence and structure, it goes on to explore the structural basis of protein function and how this function is controlled. Finally, it examines a range of case studies that illuminate the range of roles performed by proteins, and the diverse structures these proteins exhibit. Protein Structure and Function combines articulate prose with striking full colour illustrations, while the unique Primers in Biology modular structure integrates text, illustrations, definitions, and literature references for each topic into one double-page spread, bringing to the student's fingertips all the tools that they need to master that topic. Online Resource Centre The Online Resource Centre features figures from the book available to download, for registered adopters of the book.

This is the second edition of our little red book Lectins published in 1989. In the intervening years well over 10,000 articles have appeared with lectins as the main subject, and more than twice as many in which they were touched upon, as well as around 20 books. In particular, great strides have been made in several areas of lectin research, about which little was known until the late 1980s. One prominent example is animal lectins, many of which have been discovered only during the last decade and the functions of several of which have been clarified, especially as to their key role in innate immunity. Another is the structure of lectins and of their combining sites. Thus, whereas at that time the three-dimensional structures of just three lectins and a few of their complexes with sugars had been elucidated, their numbers have increased to about 160 and over 200, respectively, and continue to grow unabated. Updating the information on these and other topics resulted in a marked expansion of the book, which is now nearly four times as long as the first edition, with 226 figures and 39 tables. Still, a few topics, such as carbohydrate-binding cytokines or bacterial toxins that are sometimes considered as lectins, have been dealt with only in passing. Similarly to the first edition, Lectins II starts with an overview of the history of lectin research.

This important reference provides up-to-date information on all aspects of ribosome-inactivating proteins (RIPs). Including a list of all known RIPs, their distribution in nature, structure, genetics and chemical and immunological properties, this reference covers mechanisms of action, including the enzymatic activity on various polynucleotide substrates; the interaction with, and entry into cells; the toxicity to animals, including the pathology of poisoning; and the immunomodulatory and allergenic activity. The book further emphasizes the use of immunotoxins and other conjugates in clinical trials for the therapy of cancer and intractable pain.

Plant lectins are extensively used as tools and as bioactive proteins in different areas of biomedical and biological research. The Handbook of Plant Lectins provides a comprehensive yet concise overview of the biochemical properties, carbohydrate-binding specificity, biological activities and applications of most of the currently known plant lectins. This handbook consists of two major sections: an introductory guide and a quick reference dictionary. Part I acquaints the newcomer to the lectin field with the essential information on lectins and their importance to biomedicine:

• what are lectins?
• their carbohydrate-binding specificity
• effects on nutrition and immunology
• use in histochemistry
• application as therapeutic agents

• isolation and characterisation;
• sugar binding specificity;
• biological activities;
• applications;
• commercial availability; and,
• a bibliography.

How one goes about analyzing proteins is a constantly evolving field that is no longer solely the domain of the protein biochemist. Investi gators from diverse disciplines find themselves with the unanticipated task of identifying and analyzing a protein and studying its physical properties and biochemical interactions. In most cases, the ultimate goal remains understanding the role(s) that the target protein is playing in cellular physiology. It was my intention that this manual would make the initial steps in the discovery process less time consuming and less intimidating. This book is not meant to be read from cover to cover. The expanded Table of Contents and the index should help locate what you are seeking. My aim was to provide practically oriented information that will assist the experimentalist in benchtop problem solving. The appendices are filled with diverse information gleaned from catalogs, handbooks, and manuals that are presented in a distilled fashion designed to save trips to the library and calls to technical service representatives. The user is encouraged to expand on the tables and charts to fit individual experimental situations. This second edition pays homage to the computer explosion and the various genome projects that have revolutionized how benchtop scientific research is performed. Bioinformatics and In silica science are here to stay. However, the second edition still includes recipes for preparing buffers and methods for lysing cells."

In this book, authors who are experts in their fields describe current advances on commercial crops and key enabling technologies that will underpin future advances in biotechnology. They discuss state of the art discoveries as well as future challenges. Tremendous progress has been made in introducing novel genes and traits into plant genomes since the first creation of transgenic plants thirty years ago, and the first commercialization of genetically modified maize in 1996. Consequently, cultivation of biotech crops with useful traits has increased more than 100-fold from 1.7 million hectares in 1996 to over 175 million hectares globally in 2013. This achievement has been made possible by continued advances in understanding the basic molecular biology of regulatory sequences to modulate gene expression, enhancement of protein synthesis and new technologies for transformation of crop plants. This book has three sections that encompass knowledge on genetically modified (GM) food crops that are currently used by consumers, those that are anticipated to reach the market place in the near future and enabling technologies that will facilitate the development of next generation GM crops. Section I focuses only on genetically modified maize and soybean (3 chapters each), while Section II discusses the GM food crops rice, wheat, sorghum, vegetables and sugar cane. Section III covers exciting recent developments in several novel enabling technologies, including gene targeting, minichromosomes, and in planta transient expression systems.

Flow cytometry is a sensitive and quantitative platform for the measurement of particle fluorescence. In flow cytometry, the particles in a sample flow in single file through a focused laser beam at rates of hundreds to thousands of particles per second. During the time each particle is in the laser beam, on the order of ten microseconds, one or more fluorescent dyes associated with that particle are excited. The fluorescence emitted from each particle is collected through a microscope objective, spectrally filtered, and detected with photomultiplier tubes.
Flow cytometry is uniquely capable of the precise and quantitative molecular analysis of genomic sequence information, interactions between purified biomolecules and cellular function. Combined with automated sample handling for increased sample throughput, these features make flow cytometry a versatile platform with applications at many stages of drug discovery. Traditionally, the particles studied are cells, especially blood cells; flow cytometry is used extensively in immunology.
This volume shows how flow cytometry is integrated into modern biotechnology, dealing with issues of throughput, content, sensitivity, and high throughput informatics with applications in genomics, proteomics and protein-protein interactions, drug discovery, vaccine development, plant and reproductive biology, pharmacology and toxicology, cell-cell interactions and protein engineering.

Advances in fluorescent proteins, live-cell imaging, and superresolution instrumentation have ushered in a new era of investigations in cell biology, medicine, and physiology. From the identification of the green fluorescent protein in the jellyfish Aequorea victoria to the engineering of novel fluorescent proteins, The Fluorescent Protein Revolution explores the history, properties, and applications of these important probes. The book first traces the history of fluorescent proteins and the revolution they enabled in cellular imaging. It then discusses fluorescent proteins with novel photophysical properties. The book also covers several cutting-edge imaging applications. These include superresolution microscopy of cellular fine structures, FRET microscopy to visualize protein interactions and cell-signaling activities inside living cells, photobleaching and photoactivation techniques to visualize protein behaviors, techniques that exploit plant and algal photoreceptors to enable light-regulated control of enzymatic activities, and the noninvasive imaging of tumor-host interactions in living animals. In color throughout, this book presents the fundamental principles and latest advances in the field, including the associated development of imaging techniques that exploit fluorescent proteins. It is accessible to a broad audience, from optical imaging experts to novices needing an introduction to the field.