Monoclonal antibody against the catalytic subunit of bovine heart cAMP-dependent protein kinase and the application of enzyme purification.

A monoclonal antibody against the catalytic subunit of bovine heart cAMP-dependent protein kinase has been prepared. The antibody, M73/PK1, belongs to the heavy-chain subclass IgG3 and has a titer of 20,480. Purified M73/PK1 was obtained by ammonium sulfate precipitation and Protein A-agarose affinity chromatography and was coupled to CNBr-activated Sepharose 6MB. The catalytic subunit was purified by this affinity column. A 75% yield and 4746-fold purification were obtained. These results show that this is a suitable and effective method for the purification of the catalytic subunit of bovine heart cAMP-dependent protein kinase.