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Biomedical Sciences - Immunology | The Complement System - Methods and Protocols

The Complement System

Methods and Protocols

Series: Methods in Molecular Biology, Vol. 1100

Gadjeva, Mihaela (Ed.)

2014, XVI, 383 p. 81 illus., 27 illus. in color.

A product of Humana Press
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  • Presents current up-to-date methodologies
  • Features tips on trouble shooting and avoiding known pitfalls
  • Contains key notes and implementation advice from the experts

Complement Systems: Methods and Protocols is composed of 32 individual chapters that describe a variety of protocols  to purify and analyze the activity of the individual complement components or pathways.  It includes assays that describe detection  of complement SNPs,  clinical methods to evaluate complement system activation and data interpretation. Written in the highly successful Methods in Molecular Biologyseries 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.


Authoritative and practical, Complement Systems: Methods and Protocols provides a collection of well-established “classical” assays and recently developed “new” assays to analyze the complement system activation will be useful to a wide audience of scientists.

Content Level » Professional/practitioner

Keywords » activation in humans - activation in model animals - complement system - complement system activation - disease - health - homeostasis - immune surveillance - methodological aspects - receptors - soluble components

Related subjects » Immunology

Table of contents 

1. Overview

Mihaela Gadjeva

 2. Evaluation of Complement Function by ELISA

Anja Roos and Jorgen Wieslander

3. Classical Complement Pathway Component C1q – Purification of Human C1q, Isolation of C1q Collagen-like and Globular Head Fragments and Production of Recombinant C1q – Derivatives. Functional Characterization. 

Mihaela Kojouharova

 4. Classical Complement Pathway Components C1r and C1s – Purification from Human Serum and in Recombinant Form and Functional Characterization

Véronique Rossi, Isabelle Bally, Monique Lacroix, Gérard J Arlaud, and Nicole M Thielens

 5. Expression and Purification Methods for the Production of Recombinant Human Complement Component C2

Angela W. Norton, Paolo G.V. Martini, Lynette C. Cook , Scott Alderucci , Dianna M. Lundberg, Susan M. Fish , Charles Bedard3, John Gill,   Arthur O. Tzianabos,  and Michael F. Concino

 6. Purification and Characterisation of human and Mouse Complement C3

Marieta M Ruseva and Meike Heurich

 7. Purification of Human Complement Protein C5

Lars Sottrup-Jensen and Gregers Rom Andersen

 8. Quantification of Complement C5b-9 Binding to Cells by Flow Cytometry

Oren Moskovich and Zvi Fishelson

 9. Challenges and Opportunities in Fractionation of Recombinant Human Mannan-binding Lectin

Thomas Vorup-Jensen

 10. Genotyping of FCN and MBL2 Polymorphisms using Pyrosequencing

Lea Munthe-Fog, Hans Ole Madsen, and Peter Garred

 11. Assay for Estimation of the Functional Activity of the Mannan Binding Lectin Pathway of the Complement System

Troels R. Kjaer and Steffen Thiel

 12. Purification, Measurement of Concentration, and Functional Complement Assay of Human Ficolins

Misao Matsushita, David Kilpatrick, Hiroshi Shiraki, Yu Liu, Koichiro Tateishi, Mitsushi Tsujimura, Yuichi Endo, Teizo Fujita

 13. Purification and Functional Analysis of Human Properdin

Joseph O`Flynn, Maria C. Faber Krol, Mohamed R. Daha, Cees van Kooten

 14. Purification and Functional Characterization of C4b-binding Protein (C4BP)

Frida C. Mohlin and Anna M. Blom

 15. Purification and Functional Characterization of Factor I

Sara C. Nilsson and Anna M. Blom

 16. C1 inhibitor – Quantification and Purification

Lilian Varga  and József Dobó

 17. Purification, Quantification and Functional Analysis of Complement Factor H

Bing-Bin Yu, Beryl E. Moffatt, Marina Fedorova, Ali Ryan, Claire G.S. Villiers, James N. Arnold, Eugenie Du, Astrid Swinkels, Man Chung Li, Robert B. Sim 18. Factor H-related Proteins

Mihály Józsi and Seppo Meri

 19. Functional Evaluation of Factor H genetic and Acquired Abnormalities; Application for Atypical Hemolytic Uremic Syndrome (aHUS)

Lubka T. Roumenina,  Roxane Roquigny, Caroline Blanc, Nelly Poulain, Stephanie Ngo, Marie-Agnes Dragon-Durey, and Veronique Fremeaux-Bacchi

 20. Anti-Factor H Auto Antibodies Assay

Marie-Agnès Dragon-Durey, Caroline Blanc, Lubka Roumenina, Nelly Poulain Poulain, Stéphanie Ngo, Pauline Bordereau, Véronique Frémeaux-Bacchi

 21.  Purification of Native Surfactant Protein SP-A from Pooled Amniotic Fluid and Bronchoalveolar Lavage

Najmunisa Karbani, Eswari Dodagatta-Marri, Asif S. Qaseem, Priyaa Madhukaran, Patrick Waters, Anthony G. Tsolaki, Taruna Madan, and Uday Kishore

 22. Purification of Surfactant Protein D (SP- D) from Pooled Amniotic Fluid and Bronchoalveolar Lavage

Eswari Dodagatta-Marri, Asif Qaseem, Najmunisa Karbani, Anthony George Tsolaki, Patrick Waters, Taruna Madan, and Uday Kishore

 23. Functional Analysis of C5a Effector Responses In Vitro and In Vivo

Christian M. Karsten, Yves Laumonnier, and  Jörg Köhl

 24. Detection of Complement Receptors 1 and 2 on Mouse Splenic B Cells using Flow Cytometry

Luke R Donius and John H Weis

 25. Quantification of Complement Receptor 2 Calcium Signaling Enhancement Using Flow Cytometry

Luke R Donius and John H Weis

 26. Purification of C1q Receptors and Functional Analysis

Berhane Ghebrehiwet and Ellinor I.B. Peerschke

 27. Detection of Cell Membrane-bound CD46 using Flow Cytometry

Martin Kolev and Claudia Kemper

 28. Detection of CD59 using Flow Cytometry

Martin Kolev

 29. Functional Analysis of CD59 using Complement Dependent Cytotoxicity Assay

Martin Kolev

 30. Detection of Soluble CR3 (CD11b/CD18) by Time-resolved Immunofluorometry

Gitte Krogh Nielsen and Thomas Vorup-Jensen

 31. Analysis of the Complement Activation in Mice

Nirmal K. Banda and Kazue Takahashi

 32. Opsonophagocytic Assay

Markryan Dwyer and Mihaela Gadjeva

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