Carroll SB, Stollar BD
J Biol Chem 1983 Jan 258:24-6
Polyclonal antibodies to calf thymus RNA polymerase II were raised in laying hens. Up to 75 mg of immunoglobulin/egg yolk were extracted by the polyethylene glycol procedure of Roeder (Roeder, R.G. (1976) in RNA Polymerase (Losick, R., and Chamberlin, M., eds) pp. 285-330, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY). The concentration of specific antibody in egg yolks (IgY) was comparable to that of serum as measured by enzyme-linked immunoassay. Purified antibody was shown to be directed against enzyme by removal of enzyme activity in immune complexes precipitated by rabbit anti-chicken IgY. The antibodies recognized several of the subunits of the enzyme as determined by their reactivity with polypeptides transferred to nitrocellulose paper after gradient sodium dodecyl sulfate-gel electrophoresis. Production of antibodies in laying hens may facilitate the study of other highly conserved antigens that are poorly immunogenic in mammalian hosts.