Purification of phospholamban and characterization of its antibodies

Phospholamban was extracted from pig heart microsomal membranes with a methanol-chloroform (2:1) mixture. The proteolipid was further purified to electrophoretic homogeneity by chromatography in organic solvents and in SDS-containing medium. Purification was about 50-fold with respect to specific 32P-phospholamban radioactivity. Antisera were produced in rabbits against phospholamban. Electroblot analysis demonstrates specific binding of the antisera to purified phospholamban and to phospholamban in preparations of cardiac sarcoplasmic reticulum and sarcolemma.