| Subculturing |
Protocol: Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 to 2 X 10 exp5 viable cells/ml. Maintain cell density between 1 X 10 exp5 and 1 X 10 exp6 viable cells/ml. Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density |
| References |
Animals were immunized with the synthetic peptide IGRGNFGEVFSG(C) corresponding to residues 567 through 578 of the FES sequence (formerly residues 529 through 540 of the HC/v-fes sequence).
Only the IgG2a immunoglobulin is reactive with the antigen.
In immunoblot assays, the antibody detects an 85 kd gag-fes fusion protein encoded by the Gardner-Arnstein strain of feline sarcoma in viral transformed mink lung cells. In an ELISA test, the antibody does not cross-react with the homologous region of src, yes, erb B, mil, raf, fms, fqr, abl, mos, CDC28 or the regulatory or catalytic subunits of bovine cyclic AMP dependent protein kinase. The IgG2a and IgG2b reactivities of the antibody with the fes peptide (567 to 578) were confirmed by ELISA. IgG2a was strongly positive while IgG2b was negative.
Only the IgG2a immunoglobulin is reactive with the antigen.
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