Characterization and comparative immunoreactivity of antibody to newt (T. cristatus) globins

Rabbit antisera to newt (T. cristatus) globin were produced by repeated injections of globin and antiglobin antibodies purified by chromatography on globin-Sepharose 4B. Ouchterlony and SDS PAGE analysis indicated that the material eluted from the affinity column was rabbit IgG. The antiglobin antibodies tested by immunodiffusion and ELISA cross-reacted with native hemoglobin and globin from T. cristatus and to varying extents with globins of N. viridescens, R. pipiens and X. laevis, but not with human globin. The degree of cross-reactivity appeared to parallel the evolutionary relatedness of these species, suggesting common antigenic determinants among globins of various vertebrate species.