| 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 a synthetic polypeptide with the sequence LVADCWKKVKEERPLF corresponding to peptides 584 to 599 of the RAF-1 oncogene (formerly v-raf residues 255 to 270).
In ELISA testing, the antibody cross reacts with the homologous region of MET, sea, ros, src, SRC, yes, PKS, FMS, HIR, lck, kit. Weak activity was detected for FGR, c-mos, v-mos, MOS, erb-B, v-fms. In immunoblot testing, proteins of approximately 80 and 100 kd are detected in fms transformed 3T3 cells. The IgM and IgG2b reactivities of the antibody with the RAF-1 peptide (584 to 599) were confirmed by ELISA. IgM was strongly positive while IgG2b was negative.
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