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Production of Antibodies to Calmodulin in Rabbits and Enzyme Immunoassays for Calmodulin and Anti-Calmodulin

Production of Antibodies to Calmodulin in Rabbits and Enzyme Immunoassays for Calmodulin and Anti-Calmodulin

S Kitajima, A Seto-Ohshima, M Sano, K Kato

J Biochem. 1983 Aug;94(2):559-64.

PMID: 6355079

Abstract:

The production of anti-calmodulin antibodies in rabbit was examined with the use of performic acid-oxidized calmodulin from bovine brain or an emulsion of native calmodulin and methylated bovine serum albumin as the antigen. The antibodies in rabbit sera were determined serially by means of a sensitive enzyme immunoassay method that uses a mini-column of goat (anti-rabbit IgG) IgG-coupled Sepharose 4B for separation. The antibodies could be produced in rabbits with either antigen. Although the native calmodulin with methylated bovine serum albumin seemed to be a better antigen, the titer of antiserum was not raised to levels detectable in the double immunodiffusion test. Monospecific antibodies were purified from the antiserum, and a competitive enzyme immunoassay method for the assay of calmodulin was developed with the use of the column-separation technique employed in the assay of anti-calmodulin. The method could determine from 10 ng to 10 micrograms (0.6 pmol to 0.6 nmol) of rat calmodulin and gave no cross-reaction with S-100 protein. Calmodulin contents in rat brain determined by the present method were consistent with those measured by the bioassay using calmodulin-deficient phosphodiesterase.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42416736	Anti-phospho-Calmodulin (pThr79+Ser81) antibody produced in rabbit			Price

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