Production of polyclonal antibodies against the 40 kDa form of human 2'-5' oligoadenylate synthetase
Gribaudo, G.; Cocciolo, M.G.; Cavallo, G.; Landolfo, S.
Journal of Biological Regulators and Homeostatic Agents 1(3): 143-146
1987
ISSN/ISBN: 0393-974X PMID: 3503518 Document Number: 298233
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.