A simple and rapid method was developed to purify monospecific antibody from heterospecific antisera utilizing “protein blotting”. Proteins were separated on a polyacrylamide gel. They were then transferred to nitrocellulose paper with an electric force, and the nitrocellulose strip corresponding to the relative mobility of antigen was removed and treated with heterospecific antisera. Monospecific antibody was successively eluted with acidic buffer from the strip. By employing this technique, monospecific antibodies against rat parotid amylase, parotid deoxyribonuclease (DNase), and albumin were purified from a mixture of these antisera.
Amylase, DNase and albumin were localized in both rat parotid gland and liver by use of these purified monospecific antibodies in the immunohistochemical method.