B Cell Protocols

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.

Includes supplementary material: sn.pub/extras

Klappentext
Recent advances in genetics, synergized by many exciting developments in cellular and molecular biology, have provided powerful new experimental tools for the study of not only B-lymphocyte development and function, but also the physiology and pathology of the immune system in higher animals. In B Cell Protocols, highly qualified investigators who are either the inventors or expert users of their methods provide a diverse collection of protocols that range from the latest inventions and applications to classic, but still frequently used, methods in B cell biology. The protocols presented follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting-edge, B Cell Protocols offers both beginning and experienced researchers alike highly effective tools for exploring B-lymphocyte development and function in higher animals, as well as critical information on how best to design or modify an experimental approach that will prove productive in their own research.

Inhalt
Characterization of B Lymphopoiesis in Mouse Bone Marrow and Spleen.- Characterization of B-Cell Maturation in the Peripheral Immune System.- Identification of B-Cell Subsets.- Analysis of B-Cell Life-Span and Homeostasis.- Differentiation of B Lymphocytes From Hematopoietic Stem Cells.- Analysis of Lymphocyte Development and Function Using the RAG-Deficient Blastocyst Complementation System.- Conditional Gene Mutagenesis in B-Lineage Cells.- Analysis of the Germinal Center Reaction and In Vivo Long-Lived Plasma Cells.- T-Cell-Dependent Immune Responses, Germinal Center Development, and the Analysis of V-Gene Sequences.- of Genes Into Primary Murine Splenic B Cells Using Retrovirus Vectors.- Immunoglobulin Class Switching.- In Vitro and In Vivo Assays of B-Lymphocyte Migration.- Analysis of Antigen-Specific B-Cell Memory Directly Ex Vivo.- B-Cell Signal Transduction.- Isolation of Lipid Rafts From B Lymphocytes.- Molecular Single-Cell PCR Analysis of Rearranged Immunoglobulin Genes As a Tool to Determine the Clonal Composition of Normal and Malignant Human B Cells.- Analysis of B-Cell Immune Tolerance Induction Using Transgenic Mice.- Analysis of B-Cell Signaling Using DT40 B-Cell Line.- B-Cell Development and Pre-B-1 Cell Plasticity In Vitro.