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B-lymphocyte development and function remains an exciting area of research for those interested in the physiology and pathology of the immune system in higher animals. While recent advances in genetics and cellular and molecular biology have provided a large spectrum of powerful new experimental tools in this field, it is both time consuming and often very difficult for a student or just any bench-side worker to identify a reliable experimental protocol in the ocean of the literature. The aim of B Cell Protocols is to provide a collection of diverse protocols ranging from the latest inventions and applications to some classic, but still frequently used methods in B-cell biology. The authors of the various chapters are all highly qualified scientists who are either the inventors or expert users of these methods. Their extensive experience in mastering a particular method provides not only the step-by-step details of a reproducible protocol, but also useful troubleshooting tips that readers will appreciate in their daily work. We hope that this book will be helpful for both beginning and experienced researchers in the field in designing or modifying an experimental approach, and exploring a biological question from multiple angles.
With a wide variety of investigative approaches, T cell immunology is a vital and open field of study. In T Cell Protocols, Second Edition, an international panel of experts contribute fully updated classic protocols as well as newly established novel techniques for the study of T lymphocyte biology. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters in this volume provide brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections which collect expert tips on troubleshooting and avoiding known pitfalls. Up-to-date and easy to use, T Cell Protocols, Second Edition is an ideal guide for young investigators new to the complex field of immunology as well as a valuable, concise resource for experienced scientists searching for clear, efficacious descriptions of novel methods.
“Infogest” (Improving Health Properties of Food by Sharing our Knowledge on the Digestive Process) is an EU COST action/network in the domain of Food and Agriculture that will last for 4 years from April 4, 2011. Infogest aims at building an open international network of institutes undertaking multidisciplinary basic research on food digestion gathering scientists from different origins (food scientists, gut physiologists, nutritionists...). The network gathers 70 partners from academia, corresponding to a total of 29 countries. The three main scientific goals are: Identify the beneficial food components released in the gut during digestion; Support the effect of beneficial food components on human health; Promote harmonization of currently used digestion models Infogest meetings highlighted the need for a publication that would provide researchers with an insight into the advantages and disadvantages associated with the use of respective in vitro and ex vivo assays to evaluate the effects of foods and food bioactives on health. Such assays are particularly important in situations where a large number of foods/bioactives need to be screened rapidly and in a cost effective manner in order to ultimately identify lead foods/bioactives that can be the subject of in vivo assays. The book is an asset to researchers wishing to study the health benefits of their foods and food bioactives of interest and highlights which in vitro/ex vivo assays are of greatest relevance to their goals, what sort of outputs/data can be generated and, as noted above, highlight the strengths and weaknesses of the various assays. It is also an important resource for undergraduate students in the ‘food and health’ arena.
Current Protocols in Immunology is a three-volume looseleaf manual that provides comprehensive coverage of immunological methods from classic to the most cutting edge, including antibody detection and preparation, assays for functional activities of mouse and human cells involved in immune responses, assays for cytokines and their receptors, isolation and analysis of proteins and peptides, biochemistry of cell activation, molecular immunology, and animal models of autoimmune and inflammatory diseases. Carefully edited, step-by-step protocols replete with material lists, expert commentaries, and safety and troubleshooting tips ensure that you can duplicate the experimental results in your own laboratory. Bimonthly updates, which are filed into the looseleaf, keep the set current with the latest developments in immunology methods. The initial purchase includes one year of updates and then subscribers may renew their annual subscriptions. Current Protocols publishes a family of laboratory manuals for bioscientists, including Molecular Biology, Human Genetics, Protein Science, Cytometry, Cell Biology, Neuroscience, Pharmacology, and Toxicology.
Now completely revised and updated from the original, much-acclaimed and bestselling first edition, Basic Cell Culture Protocols, 2nd ed. offers today's most comprehensive collection of easy-to-follow, cutting-edge protocols for the culture of a wide range of animal cells. Its authoritative contributors provide explicit, step-by-step instructions, along with extensive notes and tips that allow both experts and beginners to successfully achieve their desired results. Topics range from basic culture methodology to strategies for culturing previously uncultured cell types and hard-to-culture differentiated cells. Methods are also provided for the analysis of living cells by FACS, video microscopy, and confocal microscopy. Like the first edition, this book should be in every cell culture laboratory and be of use to all who use cell cultures in research.
This volume presents up-to-date methods that allow primary stem cells from a variety of sources to be isolated, cultured in vitro, detected and measured for specific applications. These applications range from those in basic, stem cell and veterinary research to toxicology, cellular therapy and regenerative medicine. There is a slight bias towards the blood-forming system as more is known about the blood-forming or hematopoietic system than any other primary stem cell system. These unique properties and characteristics are discussed and examined, mostly at the cellular level and in detail in this book. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Stem Cell Protocols provides novices with the fundamentals necessary to develop new technologies necessary for basic and clinical research in the future, and will aid professionals in finding new methodologies to provide a wider viewpoint and an even greater scope for their own research.
This volume details our current understanding of the architecture and signaling capabilities of the B cell antigen receptor (BCR) in health and disease. The first chapters review new insights into the assembly of BCR components and their organization on the cell surface. Subsequent contributions focus on the molecular interactions that connect the BCR with major intracellular signaling pathways such as Ca2+ mobilization, membrane phospholipid metabolism, nuclear translocation of NF-kB or the activation of Bruton’s Tyrosine Kinase and MAP kinases. These elements orchestrate cytoplasmic and nuclear responses as well as cytoskeleton dynamics for antigen internalization. Furthermore, a key mechanism of how B cells remember their cognate antigen is discussed in detail. Altogether, the discoveries presented provide a better understanding of B cell biology and help to explain some B cell-mediated pathogenicities, like autoimmune phenomena or the formation of B cell tumors, while also paving the way for eventually combating these diseases.
This third edition volume expands on the previous editions with more detailed research on the characterization of antibody antigen interactions between different users with different requirements. The chapters in this book are divided into four parts: Part One looks at the entire native antigen and covers traditional structural biology techniques such as nuclear magnetic resonance and x-ray crystallography. Part Two talks about protein fragments derived from antigens, and discusses binding regions within antigen sequence using bacterial surface display and ELISA, for example. Part Three describes the use of surface plasmon resonance spectroscopy and biolayer interferometry, and Part Four highlights methods used to identify new antigens and assess antibody cross-reactivity. 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, Epitope Mapping Protocols, Third Edition is a valuable resource for anyone interested in furthering their research in this expanding field.
At some point in their careers, virtually every scientist and technician, as well as many medical professionals, regardless of their area of specialization have a need to utilize cell culture systems. Updating and significantly expanding upon the previous editions, Basic Cell Culture Protocols, Fourth Edition provides the novice cell culturist with sufficient information to perform the basic techniques, to ensure the health and identity of their cell lines, and to be able to isolate and culture specialized primary cell types. The intent of this extensive volume is to generate a valuable resource containing clear methodologies pertinent to current areas of investigation, rather than attempting to educate cell culturists on specific cell types or organ systems. Written in the highly successful Methods in Molecular BiologyTM, 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 up-to-date, Basic Cell Culture Protocols, Fourth Edition compiles the essential techniques needed to approach this vital laboratory activity with full success.
Given the versatile utility of the determinination of epitopes, beneficial to a wide variety of scientists from immunologists to structural biologists to biotechnologists, the need for a thorough, state-of-the-art collection of experimental protocols is clear. In Epitope Mapping Protocols, Second Edition, expert contributors from a broad spectrum of scientific backgrounds update and expand the successful first edition with cutting-edge techniques and applications, including approaches to both antibody or B-cell epitope mapping and T-cell epitope mapping as well as a new section on the profiling of antibody signatures in biological fluids. Written in the popular Methods in Molecular BiologyTM series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Epitope Mapping Protocols, Second Edition is a reliable and valuable reference for all those who wish to understand and further investigate the diversifying field of epitope mapping.