Affinity purification of antibodies of regulatory subunits of cAMP-dependent protein kinase using cross-linked immunoabsorbent

Abstract

The purification of antibodies of the regulatory subunit (R) of type I (R I) and type II (R II) cAMP-dependent protein kinases is achieved by using a cross-linked R I- and R II-Sepharose with 5 mM glutaraldehyde. The cross-linking eliminates the problem of ‘leaking out’ of coupled R during the affinity chromatography of R I and R II antibodies. The general utility of this method for purification of antibodies for the immunocytochemical localization of other proteins as of R is discussed.