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This volume describes a range of methods used in complement laboratories and how to interpret the data. Chapters detail methods for depletion of IgG and IgM, quantification of complement proteins, C3dg quantification, complement C3 deposition on endothelial cells, anti-C1q auto-antibodies, and methods for assessment of interactions of proteins with heme. 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.
Cutting-edge and clearly written, The Complement System: Innovative Diagnostic and Research Protocols aims to ensure successful results in the further study of this vital field.
List of contents
The Benefits of Complement Measurements for the Clinical Practice.- Method for Depletion of IgG and IgM from Human Serum as Naive Complement Source.- Quantification of Complement Proteins with Special Reference to C1q: Multiplex vs ELISA vs Rocket Immunoelectrophoresis vs Nephelometry.- C3dg Quantification by PEG-precipitation and ELISA or TRIFMA.- Quantification of Porcine Complement Activation Fragment C3a by a Neoepitope Based Enzyme-Linked Immunosorbent Assay.- Sheep Erythrocytes Preparation for Hemolytic Tests Exploring Complement Functional Activities.- Hemolytic Tests Exploring Factor H Functional Activities.- Functional Hemolytic Test for Complement Alternative Pathway Convertase Activity.- Complement C3 Deposition on Endothelial Cells Revealed by Flow Cytometry.- Anti-C1q Auto-Antibodies: Standard Quantification and Innovative ELISA.- Anti-C1-Inhibitor Auto-Antibodies Detection by ELISA.- Anti-ficolin-2 and anti-ficolin-3 Autoantibodies Detection by ELISA.- Detection of anti-C3b Autoantibodies by ELISA.- Detection of Complement Factor B Autoantibodies by ELISA.- Detection of C3 Nephritic Factor by Hemolytic Assay.- Detection of Genetic Rearrangements inthe Regulators of Complement Activation RCA Cluster by High Throughput Sequencing and MLPA.- Complement Detection in Mouse Kidneys by Immunofluorescence.- Complement Detection in Human Tumors by Immunohistochemistry and Immunofluorescence.- Analysis Of The Ligand Recognition Specificities Of Human Ficolins Using Surface Plasmon Resonance.- Methods for Assessment of Interactions of Proteins with Heme - Application for Complement Proteins and Immunoglobulins.- Evaluation of Binding Kinetics and Thermodynamics of Antibody-antigen Interactions and Interactions Involving Complement Proteins.- Purification of Human Complement Component C4 and Sample Preparation for Structural Biology Applications.
