Production of polyclonal antibodies against the 40 kDa form of human 2'-5' oligoadenylate synthetase.
A 17-aminoacid peptide corresponding to the C terminal of the smaller form of human 2'-5' oligoadenylate synthetase was coupled to keyole lymphet haemocyanin (KLH) and used as immunogen in rabbits. After a cycle of four immunizations two animals produced immunoglobulins able to recognize the 17-aminoacid peptide as evaluated in ELISA assays. The specific Ig were purified by an immunoadsorbent with the peptide immobilized on Sepharose CL-4B and used in Western blot employing either protein A iodinated or conjugated with peroxidase as indicator system. The results obtained using extracts from HeLa or WISH cells treated for 15 hr with HuIFN-alfa as antigen demonstrate that the anti-peptide antibodies recognize the 40 kDa form of the 2'-5' oligoadenylate synthetase enzyme complex. These antibodies therefore represent a useful tool for monitoring the induction of the above enzyme.
D015088:2',5'-Oligoadenylate Synthetase / Q000096:biosynthesis / Q000276:immunology*; D000917:Antibody Formation; D015153:Blotting, Western; D004790:Enzyme Induction; D006367:HeLa Cells; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000494:pharmacology; D008970:Molecular Weight; D011994:Recombinant Proteins; D014407:Tumor Cells, Cultured
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D015088:2',5'-Oligoadenylate Synthetase