ANTIBODIES RAISED AGAINST SYNTHETIC PEPTIDES REACT WITH CHOLINE-ACETYLTRANSFERASE IN VARIOUS IMMUNOASSAYS AND IN IMMUNOHISTOCHEMISTRY

Antisera were raised in rabbits against five synthetic peptides. These peptides have been identified as potentially antigenic epitopes from the sequence of porcine choline acetyltransferase (ChAT) using primary and secondary structure analysis. All five antisera recognized immuno affinity-purified antigen from porcine brain in an ELISA and on wester n blots. Four antisera recognized ChAT on dot blots, and another four antisera reacted with native and degraded enzyme in a sandwich ELISA u sing monoclonal antibodies as the capture antibody. One peptide antise rum was of similar avidity in this sandwich ELISA as a polyclonal anti body raised against immunoaffinity-purified ChAT. The same antiserum r eacted with the enzyme from human placenta in an ELISA and on western and dot blots and recognized ChAT in rat, primate, and human neurons. Thus, a single peptide (amino acids 168-189) provides the means for ea sy, reliable, and reproducible generation of antibodies against ChAT s uitable for replacing conventional polyclonal and monoclonal antibodie s.