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.

Characterization of B Lymphopoiesis in Mouse Bone Marrow and Spleen Richard R. Hardy and Susan A. Shinton Characterization of B-Cell Maturation in the Peripheral Immune System Rita Carsetti Identification of B-Cell Subsets: An Exposition of 11-Color (Hi-D) FACS Methods James W. Tung, David R. Parks, Wayne A. Moore, Leonard A. Herzenberg, and Leonore A. Herzenberg Analysis of B-Cell Life-Span and Homeostasis Irmgard Förster Differentiation of B Lymphocytes From Hematopoietic Stem Cells Paulo Vieira and Ana Cumano Analysis of Lymphocyte Development and Function Using the RAG-Deficient Blastocyst Complementation System Faith M. Young, Carl A. Pinkert, and Andrea Bottaro Conditional Gene Mutagenesis in B-Lineage Cells Stefano Casola Analysis of the Germinal Center Reaction and In Vivo Long-Lived Plasma Cells Kai-Michael Toellner, Mahmood Khan, and Daniel Man-Yuen Sze T-Cell-Dependent Immune Responses, Germinal Center Development, and the Anaysis of V-Gene Sequences Claudia Berek Introduction of Genes Into Primary Murine Splenic B Cells Using Retrovirus Vectors Kuo-I Lin and Kathryn Calame Immunoglobulin Class Switching: In Vitro Induction and Analysis Sven Kracker and Andreas Radbruch In Vitro and In Vivo Assays of B-Lymphocyte Migration Chantal Moratz and John H. Kehrl Analysis of Antigen-Specific B-Cell Memory Directly Ex Vivo Louise J. McHeyzer-Williams and Michael G. McHeyzer-Williams B-Cell Signal Transduction: Tyrosine Phosphorylation, Kinase Activity, and Calcium Mobilization Tilman Brummer, Winfried Elis, Michael Reth, and Michael Huber Isolation of Lipid Rafts From B Lymphocytes Anu Cherukuri, Shiang-Jong Tzeng, Arun Gidwani, Hae Won Sohn, Pavel Tolar, Michelle D. Snyder, and Susan K. Pierce Molecular Single-Cell PCR Analysis of Rearranged Immunoglobulin Genes As a Tool to Determine the Clonal Composition of Normal and Malignant Human B Cells Ralf Küppers Analysis of B-Cell Immune Tolerance Induction Using Transgenic Mice Helen Ferry and Richard J. Cornall Analysis of B-Cell Signaling Using DT40 B-Cell Line Tomoharu Yasuda and Tadashi Yamamoto B-Cell Development and Pre-B-1 Cell Plasticity In Vitro Antonius G. Rolink Index