Preparation of a monoclonal antibody against rat MnSOD, using a COOH-terminal peptide.
In the present paper we report the production of a monoclonal antibody against rat MnSOD, a supposed tumor-suppressor protein, using a purified synthetic peptide encompassing amino acids 184-198 to immunize mice, without conjugation to a carrier. The resulting antibody is able to recognize the native form of the protein, since it can immunoprecipitate the MnSOD activity in rat liver homogenate. In Western blot studies, the antibody recognizes a protein of 24 KD M(r), whose concentration varies according to the MnSOD activity and it apparently recognizes also human and mouse MnSODs. The protocol of immunization gives high yield of secreting lines. This monoclonal antibody will allow the detection of structural and functional alterations of MnSOD.
Ria
F
F
Institute of General Pathology, Catholic University, Rome, Italy.
Landriscina
M
M
Galeotti
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006801:Humans; D006825:Hybridomas; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008930:Mitochondria, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology; D051381:Rats; D011912:Rats, Inbred ACI; D011915:Rats, Inbred BUF; D017208:Rats, Wistar; D013482:Superoxide Dismutase / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009842:Oligopeptides; D013482:Superoxide Dismutase