Complement Methods and Protocols

The complement system, first described more than a century ago, was for many years the ugly duckling of the immunology world, but no more. Complement in recent years has blossomed into a fascinating and fast moving field of immediate relevance to clinical scientists in fields as diverse as transplantation biology, virology, and inflammation. Despite its emergence from the shadows, complement retains an unwarranted reputation for being difficult. This impression derives in large part from the superficially complicated nomenclature, a relic of the long and tortuous process of unraveling the system, of naming components in order of discovery rather than in a syst- atic manner. Once the barrier of nomenclature has been surmounted, then the true simplicity of the system becomes apparent. Complement comprises an activation system and a cytolytic system. The former has diverged to focus on complement to distinct targetsbacteria, - mune complexes, and othersso that texts now describe three activation pa- ways, closely related to one another, but each with some unique features. The cytolytic pathway is the same regardless of the activation process and kills cells by creating pores in the membrane. Complement plays an important role in killing bacteria and is essential for the proper handling of immune complexes. Problems occur when complement is activated in an inappropriate mannerthe potent inflammation-inducing products of the cascade then cause unwanted tissue damage and destruction.

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Klappentext

B. Paul Morgan and a team of expert laboratorians present a comprehensive set of readily reproducible methods to study this critical system. These cutting-edge techniques are suitable both for the basic scientist interested in understanding complement's mechanisms of activation and for the clinical scientist wishing to quantify its activation, and range from the purification of its components to generating complement-deficient mice by gene deletion. Additional techniques presented include procedures for the analysis of its function, for the study of its regulators, for detection of its activation in vivo, and for the identification of its autoantibodies. Comprehensive and cutting-edge, Complement Methods and Protocols offers today's basic and clinical investigators powerful tools for the analysis of the role of complement in human pathophysiology and disease, as well as its therapeutic regulation.

Inhalt
The Complement System:An Overview.- Purification of Complement Components, Regulators, and Receptors by Classical Methods.- Immunoaffinity Methods for Purification of Complement Components and Regulators.- Measurement of Complement Hemolytic Activity, Generation of Complement-Depleted Sera, and Production of Hemolytic Intermediates.- Measurement of Complement Lysis of Nucleated Cells.- Functional Assays for Complement Regulators.- Immunochemical Measurement of Complement Components and Activation Products.- Complement Deposition in Tissues.- Complement Regulators and Receptors in Tissues.- Measurement of C3 Fragment Deposition on Cells.- Screening for Complement Deficiency.- C1-Inhibitor: Antigenic and Functional Analysis.- Autoantibodies to Complement Components.- Allotyping of Complement Components.- Complement and Immune Complexes.- Knocking Out Complement Genes.- Inherited Complement Deficiencies in Animals.