Main description:

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.

Contents:

1. The Benefits of Complement Measurements for the Clinical Practice

Anne Grunenwald and Lubka T. Roumenina

2. Method for Depletion of IgG and IgM from Human Serum as Naive Complement Source

Seline A. Zwarthoff, Simone Magnoni, Piet C. Aerts, Kok P. M. van Kessel, and Suzan H.M. Rooijakkers

3. Quantification of Complement Proteins with Special Reference to C1q: Multiplex vs ELISA vs Rocket Immunoelectrophoresis vs Nephelometry

Kerstin Sandholm, Barbro Persson, Sliva Abdalla, Camilla Mohlin, Bo Nilsson, and Kristina N Ekdahl

4. C3dg Quantification by PEG-precipitation and ELISA or TRIFMA

Anne Troldborg and Jens Christian Jensenius

5. Quantification of Porcine Complement Activation Fragment C3a by a Neoepitope Based Enzyme-Linked Immunosorbent Assay

Per H. Nilsson, Kristin Pettersen, Martin Oppermann, Espen Skjeflo, Hilde Fure, Dorte Christiansen, and Tom Eirik Mollnes

6. Sheep Erythrocytes Preparation for Hemolytic Tests Exploring Complement Functional Activities

Melchior Chabannes, Pauline Bordereau, Paula Vieira Martins, and Marie-Agnes Dragon-Durey

7. Hemolytic Tests Exploring Factor H Functional Activities

Melchior Chabannes, Shambhuprasad K Togarsimalemath, and Marie-Agnes Dragon-Durey

8. Functional Hemolytic Test for Complement Alternative Pathway Convertase Activity

Marloes A.H.M. Michels, Nicole C.A.J. van de Kar, Elena B. Volokhin, and Bert (L) P.W.J. van den Heuvel

9. Complement C3 Deposition on Endothelial Cells Revealed by Flow Cytometry

Idris Boudhabhay, Anne Grunenwald, and Lubka T. Roumenina

10. Anti-C1q Auto-Antibodies: Standard Quantification and Innovative ELISA

Kinga Csorba, Lucias Schirmbeck, Denise Dubler, and Marten Trendelenburg

11. Anti-C1-Inhibitor Auto-Antibodies Detection by ELISA

Chiara Suffritti, Sonia Caccia, Silvia Berra, Debora Parolin, and Marco Cicardi

12. Anti-ficolin-2 and anti-ficolin-3 Autoantibodies Detection by ELISA

Chantal Dumestre-Perard and Nicole M. Thielens

13. Detection of anti-C3b Autoantibodies by ELISA

Maria Radanova, Lubka Roumenina, and Vasil Vasilev

14. Detection of Complement Factor B Autoantibodies by ELISA

Mihaly Jozsi and Barbara Uzonyi

15. Detection of C3 Nephritic Factor by Hemolytic Assay

Melchior Chabannes, Veronique Fremeaux-Bacchi, Sophie Chauvet

16. Detection of Genetic Rearrangements inthe Regulators of Complement Activation RCA Cluster by High Throughput Sequencing and MLPA

Jesus Garcia-Fernandez, Susana Vilches-Arroyo, Leticia Olavarrieta, Julian Perez-Perez, and Santiago Rodriguez de Cordoba

17. Complement Detection in Mouse Kidneys by Immunofluorescence

Jennifer Laskowski and Joshua M. Thurman

18. Complement Detection in Human Tumors by Immunohistochemistry and Immunofluorescence

Marie V. Daugan, Margot Revel, Laetitia Lacroix, Catherine Sautes-Fridman, Wolf H. Fridman, and Lubka T. Roumenina

19. Analysis Of The Ligand Recognition Specificities Of Human Ficolins Using Surface Plasmon Resonance

Nicole M. Thielens, Evelyne Gout, Monique Lacroix, Jean-Baptiste Reiser, and Christine Gaboriaud

20. Methods for Assessment of Interactions of Proteins with Heme - Application for Complement Proteins and Immunoglobulins

Margot Revel and Jordan D. Dimitrov

21. Evaluation of Binding Kinetics and Thermodynamics of Antibody-antigen Interactions and Interactions Involving Complement Proteins

Sofia Rossini and Jordan D. Dimitrov

22. Purification of Human Complement Component C4 and Sample Preparation for Structural Biology Applications

Alessandra Zarantonello, Sofia Mortensen, Nick S. Laursen, and Gregers R. Andersen