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Generation of polyclonal antibodies against multiple proteins in a single rabbit and subsequent isolation of the specific immunoglobulins as a tool for proteomics research
Author(s) -
Jacquot JeanPierre,
Rouhier Nicolas,
Navrot Nicolas,
Decottignies JeanPierre
Publication year - 2004
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20030196
Subject(s) - polyclonal antibodies , thioredoxin , antibody , western blot , microbiology and biotechnology , biochemistry , antigen , chemistry , sepharose , biology , enzyme , immunology , gene
A single rabbit has been immunized with a mixture of six different proteins: green fluorescent protein, ferredoxin:NADP oxidoreductase, NADPH:thioredoxin reductase, thioredoxin f , fructose bisphosphatase and cyclophilin. Individual IgGs were re‐purified from the serum through specific interaction on small columns generated by grafting the antigen on a CNBr‐Sepharose matrix. The specificity of the isolated IgG was checked by Western‐blot analysis. Purified and specific IgG fractions were obtained with all antigens except thioredoxin f .