Preparation of antisera to the mu chain of IgM.

The protein fraction in the first peak obtained by Sephadex G-200 gel filtration of normal bovine serum was reduced with 2-mercaptoethanol, alkylated with iodoactamide and recycled on a Sephadex G-200 column. The resultant chromatogram indicated the presence of two peaks the second of which was found to be pure monomeric IgM. Immunization of rabbits with the second peak protein resulted in the production of an antiserum with both heavy and light-chain activity. The light-chain activity was simply removed by absorption of the antiserum with glutaraldehyde-polymerized material obtained from the second peak of the initial Sephadex G-200 eluate, yielding an antiserum monospecific for the heavy chain of the IgM molecule.