Preparation of rat monoclonal antibodies to epitopes encoded by the viral oncogene (v-fms) of McDonough feline sarcoma virus
✍ Scribed by Soni J. Anderson; Mark E. Furth; Linda Wolff; Sandra K. Ruscetti; Charles J. Sherr
- Publisher
- John Wiley and Sons
- Year
- 1982
- Tongue
- English
- Weight
- 359 KB
- Volume
- 19
- Category
- Article
- ISSN
- 0730-2312
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✦ Synopsis
Abstract
The McDonough strain of feline sarcoma virus (SM‐FeSV) contains a viral oncogene, v‐fms, transduced from cat cellular genetic sequences designated c‐fms. Monoclonal antibodies reactive to antigenic determinants encoded by v‐fms were prepared by immunizing rats with live, syngeneic SM‐FeSV‐transformed cells, and fusing splenic lymphocytes from a tumor‐bearing animal with cultured rat myeloma cells. Culture supernatants from hybrids producing antibodies to epitopes encoded by v‐fms were identified by immunoprecipitation of radiolabeled polypeptides from SM‐FeSV‐transformed mink cells. Four positive hybrids were cloned twice in soft agar, established as stable lines, and grown in defined serum‐free medium to facilitate purification of homogeneous antibodies. The monoclonal antibodies were used to assay SM‐FeSV‐specific products by “immunoblotting” of elcctrophoretically separated proteins, and by fixed‐cell immunofluorescence.