Absorption of antisera for studies on specific enzyme turnover

Abstract

Antisera [sheep] were raised to acetyl-CoA carboxylase and 6-phosphogluconate dehydrogenase from mammary glands of lactating rabbits and to cytochrome oxidase from rat liver. The enzymes were all highly purified but gave rise to multispecific antisera when tested against tissue extracts. Absorption procedures were devised to free the antisera of contaminating antibodies. Antisera to acetyl-CoA carboxylase and cytochrome oxidase were absorbed with fractions discarded during enzyme purification. The antiserum to 6-phosphogluconate dehydrogenase was absorbed with a tissue extract from an early stage in mammary-gland differentiation. Monospecific antisera are essential for enzyme turnover studies, and therefore antisera should be extensively tested and absorbed before use. A general procedure for the absorption of antisera to purified enzymes was devised on the basis of accepted principles of antisera absorption.