133-10F6 (ATCC® CRL-2738)

Organism: Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)  /  Cell Type: hybridoma: B lymphocyte  / 

Organism Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
Cell Type hybridoma: B lymphocyte
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
Applications
Using immunoblot techniques, the ascites produced from these cells recognized proteins of approximately 45,000 and 75,000 daltons in a variety of transformed mammalian cells.
In addition, proteins of 45,000 and 55,000 were detected in various urine samples.
Animals were immunized with a synthetic polypeptide with the sequence LGEHHCTPSPPVDHG corresponding to peptides 348 to 363 of the c-myb oncogene, formerly v-myb (residues 160 to 175).
Storage Conditions liquid nitrogen vapor phase
Derivation
Animals were immunized with a synthetic polypeptide with the sequence LGEHHCTPSPPVDHG corresponding to peptides 348 to 363 of the c-myb oncogene, formerly v-myb (residues 160 to 175). Spleen cells were fused with Sp2/0 myeloma cells. Using immunoblot techniques, the ascites produced from these cells recognized proteins of approximately 45,000 and 75,000 daltons in a variety of transformed mammalian cells. In addition, proteins of 45,000 and 55,000 were detected in various urine samples. The IgG1 and IgG2b reactivities of the antibody with the c-myb peptide (348 to 363) were confirmed by ELISA. IgG1 was strongly positive while IgG2b was only slightly reactive. A culture submitted to the ATCC in February of 2001 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.
Genes Expressed
immunoglobulin; monoclonal antibody; against a synthetic c-myb oncogene peptide
Cellular Products
immunoglobulin; monoclonal antibody; against a synthetic c-myb oncogene peptide
Tumorigenic Yes
Effects
Yes, produces ascites in female 129G1X+ x BALB/c mice
Comments
Animals were immunized with a synthetic polypeptide with the sequence LGEHHCTPSPPVDHG corresponding to peptides 348 to 363 of the c-myb oncogene, formerly v-myb (residues 160 to 175). Spleen cells were fused with Sp2/0 myeloma cells. Using immunoblot techniques, the ascites produced from these cells recognized proteins of approximately 45,000 and 75,000 daltons in a variety of transformed mammalian cells. In addition, proteins of 45,000 and 55,000 were detected in various urine samples. The IgG1 and IgG2b reactivities of the antibody with the c-myb peptide (348 to 363) were confirmed by ELISA. IgG1 was strongly positive while IgG2b was only slightly reactive. A culture submitted to the ATCC in February of 2001 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
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(5) viable cells/ml.
Interval: Maintain cell density between 1 X 10(5) and 1 X 10(6) viable cells/ml.
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Isotype IgG1 and 2b kappa
Name of Depositor National Cancer Institute