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A simple procedure for isolation of Bufo arenarum C3 complement fraction and preparation of antiserum.

Authors
  • Llanos, R J
  • Miceli, D C
  • Valz-Gianinet, J N
  • Whitacre, C M
Type
Published Article
Journal
Comparative biochemistry and physiology. Part C, Pharmacology, toxicology & endocrinology
Publication Date
Sep 01, 1999
Volume
124
Issue
1
Pages
1–5
Identifiers
PMID: 10579641
Source
Medline
License
Unknown

Abstract

Because of the need for antibodies in our studies involving the third component of complement in Bufo arenarum, we performed a simple procedure to purify C3 from B. arenarum serum to use as antigen in the preparation of the antiserum. The strategy was based on the well-known ability of C3 to bind to zymosan (Zy), a yeast cell wall extract comprised of polysaccharides. The Zy-bound fraction showed cross reactivity with a commercial antibody to human C3 as well as a similar electrophoretic profile (SDS-PAGE) to C3 from other species. The Zy-C3 complex resulting from binding Zy to B. arenarum serum was injected into rabbits and the antiserum against this C3-like fraction was purified by protein A-Sepharose chromatography. The purified C3 antibody was found to be suitable for immunochemical studies.

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