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Updated: Jul 19, 2026
Pull-down of Calmodulin-binding Proteins
Published on: January 23, 2012
Production of antibody against calmodulin which is a protein with poor antigenicity
1Department of Microbiology, School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
Abstract:
Calmodulin (CaM), a Ca(2+)-binding protein, is a multifunctional modulator mediating the calcium regulation of a large number of intracellular enzyme systems. Because of its highly conserved 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.
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