PRODUCTION OF MONOCLONAL CYCLIC-GMP AND CYCLIC-AMP ANTISERA

  • 1 January 1985
    • journal article
    • research article
    • Vol. 10  (2) , 189-196
Abstract
To facilitate the in situ localization of cyclic nucleotides, a large supply of high-titer anti-sera for immunohistologic analyses would be valuable. Monoclonal antibodies (Mab) directed at 2''-0-succinyl cGMP (s-cGMP) or 2''-0-succinyl-cAMP (s-cAMP) were generated. Immunized mice developed polyclonal antibodies of high titer to either cyclic nucleotide as reflected in an enzyme-linked immunosorbent assay (ELISA) employed to monitor these anti-sera. With the use of standard hybridoma technology and ELISA to monitor anti-cyclic nucleotides antibody production from positive clones, several Mab-producing clones were grown to large volume in tissue culture and ascites fluid. The anti-cGMP and anti-cAMP anti-sera, which demonstrated significant ELISA titers at greater than 1:2,000,000 dilutions, are monospecific and can be employed for radioimmunoassays, ELISA or immunohistologic localization of cyclic nucleotides in situ.