Production of antibodies to calmodulin in rabbits and enzyme immunoassays for calmodulin and anti-calmodulin.
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.
D000818:Animals; D000906:Antibodies / Q000032:analysis; D000917:Antibody Formation; D002147:Calmodulin / Q000032:analysis / Q000276:immunology*; D002417:Cattle; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000302:isolation & purification; D008297:Male; D011817:Rabbits; D051381:Rats
D000906:Antibodies; D002147:Calmodulin; D007074:Immunoglobulin G