Production of sheep polyclonal antibodies against the 2'-5' oligoadenylate synthetases.
A 20-amino acid peptide corresponding to a common part of the 40 and 46 kD forms of human 2'-5' oligoadenylate synthetase was coupled to keyole lymphet haemocyanin (KLH) and used as immunogen in sheep. After a cycle of four immunizations, immunoglobulins able to recognize the 20-amino acid peptide as evaluated in ELISA assays were purified by an immunoadsorbent with the peptide immobilized on Sepharose CL-4B and used in Western blot employing a secondary anti-sheep antibodies and iodinated protein A as indicator system. Results obtained using extracts from human and mouse cells treated for 15 hr with IFN-alpha as antigen demonstrated that the anti-peptide antibodies recognize several forms of the 2'-5' oligoadenylate synthetase enzyme complex. These antibodies therefore represent a useful tool for monitoring the induction of the above enzymes.
Gribaudo
G
G
Istituto di Microbiologia, Università degli Studi di Torino, Italy.
Franco
A
A
Gariglio
M
M
Martinotti
M G
MG
D015088:2',5'-Oligoadenylate Synthetase / Q000096:biosynthesis / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004790:Enzyme Induction / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology; D006367:HeLa Cells / Q000187:drug effects / Q000201:enzymology; D006433:Hemocyanins / Q000276:immunology; D006801:Humans; D007163:Immunosorbent Techniques; D016898:Interferon-alpha / Q000494:pharmacology; D007518:Isoantibodies / Q000276:immunology* / Q000302:isolation & purification; D007739:L Cells / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D012756:Sheep / Q000276:immunology
D016898:Interferon-alpha; D007518:Isoantibodies; D010446:Peptide Fragments; D006433:Hemocyanins; D015088:2',5'-Oligoadenylate Synthetase; C032808:keyhole-limpet hemocyanin