Production of Antibody against Calmodulin Which Is a Protein with Poor Antigenicity

Abstract

Calmodulin (CaM), a Ca²⁺-binding protein, is a multifunctional modulator mediating the calcium regulation of a large number of intracellular enzyme systems. Because of its highly concerved structure, it has been difficult to produce anti-CaM antibody. We describe here a simplified method for production of this antibody. BALB/c mice were immunized with a polymerized CaM, and the serum was prepared and tested for the presence of anti-CaM antibody by immunoblot analysis. The serum obtained from the immunized mice was shown to contain anti-CaM antibody. Immunoblot analysis proved that a 500-fold diluted serum was able to recognize both types of CaM with a polymerized form and monomer form. The method developed in this study may be applicable to a wide range of proteins for enhancement of antigenicity.