Generation of polyclonal antibodies against multiple proteins in a single rabbit and subsequent isolation of the specific immunoglobulins as a tool for proteomics research.
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.
Jacquot
Jean-Pierre
JP
UMR INRA UHP 1136, Interaction Arbres Microorganismes, Université Henri Poincaré, Faculté des Sciences, BP 239, 54506 Vandoeuvre Cédex, France. j2p@scbiol.uhp-nancy.fr
Rouhier
Nicolas
N
Navrot
Nicolas
N
Decottignies
Jean-Pierre
JP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification*; D000917:Antibody Formation / Q000502:physiology*; D002846:Chromatography, Affinity / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology* / Q000302:isolation & purification*; D011817:Rabbits
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G