Immunogen: Synthetic peptide RKGSSSNEPSSDSLSSPTLL(C) (residues 359-378 of the FOS sequence) coupled to keyhole limpet hemocyanin Source and Sex of Animals: Male 129GIX+ mice were used for immunization. Fusion Partners: 1.3 x 10(8) nucleated spleen cells with 2 x 10(7) SP2/0 non-secreting myeloma cells Cross-reactivity: In immunoblot screening a protein of approximately 55 kd is detected in a fibroblast cell line transformed by fes, sis, and scr. Immunization Schedule: Approximately 70 mg of coupled peptide in 300 ml of a 1:1 mixture of phosphate buffered saline pH 7.4 and complete Freund's adjuvant administered intraperitoneally on day 0; followed by an equal amount of conjugate in 300 ml of a 1:1 mixture of phosphate buffered saline pH 7.4 and 10 mg/ml alum administered intraperitoneally on day 14; followed by 70 mg of coupled peptide in 200 ml of phosphate buffered saline pH 7.4 administered intravenously on day 51. The spleen was removed on day 56. Preparation: Monoclonal antibody is in ascites fluid collected from 2-06-87 to 7-06-87. The ascites fluid was ammonium sulfate precipitated, reconstituted, filter sterilized, aliquoted 1 ml per vial, and lyophilized. The sterility is not guaranteed. Resuspend in 1 ml distilled water. Approximately 1:100,000 against the unconjugated synthetic peptide in an indirect enzyme-linked immunosorbent assay Vials may be stored at 2-8°C. IgG1 |
