title: A simple procedure for isolation of Bufo arenarum C3 complement fraction and preparation of antiserum.
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.
authors.affiliation: Department of Developmental Biology, INSIBIO (CONICET-UNT), Chacabuco, Tucuman, Argentina.
authors.forename: R J
authors.initials: RJ
authors.lastname: Llanos
authors.forename: D C
authors.initials: DC
authors.lastname: Miceli
authors.forename: J N
authors.initials: JN
authors.lastname: Valz-Gianinet
authors.forename: C M
authors.initials: CM
authors.lastname: Whitacre
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D002025:Bufonidae / Q000276:immunology*; D005591:Chemical Fractionation; D003176:Complement C3 / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D003179:Complement C3b / Q000737:chemistry / Q000302:isolation & purification; D003429:Cross Reactions; D006801:Humans; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification; D015151:Immunoblotting; D016056:Immunodominant Epitopes; D011817:Rabbits; D015054:Zymosan / Q000276:immunology
chemical_list: D003176:Complement C3; D007106:Immune Sera; D016056:Immunodominant Epitopes; D003179:Complement C3b; D015054:Zymosan