Monoclonal antibodies (McAb) were prepared by fusing sp2/0 myeoma cell with spleen cell of mice Balb/c which were immunized with 64 kDa rabbit oviductin (DPF-1). Supernatants of hybridoma cell were screened for produc...Monoclonal antibodies (McAb) were prepared by fusing sp2/0 myeoma cell with spleen cell of mice Balb/c which were immunized with 64 kDa rabbit oviductin (DPF-1). Supernatants of hybridoma cell were screened for producing anti-DPF1 McAbs with enzyme-linked immunosorbent assay (ELISA) and Western blotting. Positive hybrids were cloned by using the technique of limiting dilution. Two strains of hybridoma cell were obtained,named 3E8 and 5A7. Both McAbs were determined as I'm. Results of Western blotting showed that the two McAbs recognised the DPF-1 hand (64kDa) specifically. By using McAb-5A7 as probe, we found that DPF-1 did not occur in the tissues of spleen, uterus, ovary,small intestine, skeleton muscle, brain, liver,heart, lung and kidney, except for that of oviduct; some DPF-1 homologous molecules were also revealed itself in that of oviduct tissues of mouse and golden hamster Their apparent molecular weights were 32kDa,72kDa in mouse, and 49kDa in golden hamster. All these results indicated that DPF-1 is tissue specific not species specific. (To whon all corrcspondcnce should be addrcssed. Prcsent addrcss Shanghai Institute of Planned Parenthood Rcscarch. Shanghai 200032. PRC. )展开更多
文摘Monoclonal antibodies (McAb) were prepared by fusing sp2/0 myeoma cell with spleen cell of mice Balb/c which were immunized with 64 kDa rabbit oviductin (DPF-1). Supernatants of hybridoma cell were screened for producing anti-DPF1 McAbs with enzyme-linked immunosorbent assay (ELISA) and Western blotting. Positive hybrids were cloned by using the technique of limiting dilution. Two strains of hybridoma cell were obtained,named 3E8 and 5A7. Both McAbs were determined as I'm. Results of Western blotting showed that the two McAbs recognised the DPF-1 hand (64kDa) specifically. By using McAb-5A7 as probe, we found that DPF-1 did not occur in the tissues of spleen, uterus, ovary,small intestine, skeleton muscle, brain, liver,heart, lung and kidney, except for that of oviduct; some DPF-1 homologous molecules were also revealed itself in that of oviduct tissues of mouse and golden hamster Their apparent molecular weights were 32kDa,72kDa in mouse, and 49kDa in golden hamster. All these results indicated that DPF-1 is tissue specific not species specific. (To whon all corrcspondcnce should be addrcssed. Prcsent addrcss Shanghai Institute of Planned Parenthood Rcscarch. Shanghai 200032. PRC. )
