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Open Access Methodology article

The complement system of the goat: Haemolytic assays and isolation of major proteins

Isabel Moreno-Indias12, Alister W Dodds2, Anastasio Argüello1*, Noemi Castro1 and Robert B Sim2

Author affiliations

1 Animal Science Department, Universidad de las Palmas de Gran Canaria, Arucas, Las Palmas, 35413, Spain

2 MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK

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Citation and License

BMC Veterinary Research 2012, 8:91  doi:10.1186/1746-6148-8-91

Published: 26 June 2012

Abstract

Background

The aim of the present study was to develop a haemolytic assay for the study of the complement system in dairy goats (Capra aegagrus hircus) and to characterize the major goat complement system proteins.

Results

The commonly used sheep erythrocyte sensitized with rabbit antibodies were not sensitive to lysis by goat serum, but the combination of human red blood cells (RBC) plus rabbit antibodies was the best option found for goat complement assay. A buffer based on HEPES instead of the classical veronal (barbitone) was developed. Three proteins were isolated: factor H, C1q and C3 and these were compared with the corresponding human proteins. A novel affinity chromatography technique was developed for isolation of factor H.

Conclusions

Human RBC plus rabbit antibodies were a suitable option for haemolytic assays. The isolated proteins are similar to the human counterparts.

Keywords:
Goat; Complement system; C3; C1q; Factor H