Aw3.18.14 (ATCC® CRL-2826)

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

Organism Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
Tissue spleen
Cell Type hybridoma: B lymphocyte; somatic cell hybrid
Product Format frozen
Morphology lymphoblast
Culture Properties suspension, with some loosely adherent cells
Biosafety Level 1
Strain CAF1
Storage Conditions liquid nitrogen vapor phase
Images
Derivation

Animals were immunized with M12-I-Ak 48-62 cells. RefDadaglio G, et al. Characterization and quantitation of peptide-MHC complexes produced from hen egg lysozyme using a monoclonal antibody. Immunity 6: 727-738, 1997. PubMed: 9208845

Spleen cells were fused with P3X63Ag8 myeloma cells. 

Comments
The antibody recognizes peptide residues 48-62 of hen egg lysozyme (HEL) bound to the MHC class II molecule I-Ak.
Complete Growth Medium Dulbecco's Modified Eagle's Medium with 4 mM L-glutamine that is modified by ATCC to contain 4.5 g/L glucose and 1.5 g/L sodium bicarbonates supplemented with 1.5 mM L-glutamine, 0.05 mM 2-mercaptoethanol, 10 mM HEPES, 116mg/L L-arginine HCl, 41 mg/L L-asparagine.H2O and 10% heat-inactivated fetal bovine serum
Subculturing
Cultures can be maintained by the addition of fresh medium or replacement of medium. Shake off the attached cells and transfer along with the floating cells into new flasks. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 to 3 x 104 viable cells/mL. Maintain cell density between 2 x 104 and 2 x 105 viable cells/mL.
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density).
Cryopreservation
Complete growth medium supplemented with an additional 10% heat-inactivated fetal bovine serum and 7.5% DMSO. Cell culture tested DMSO is available as ATCC® Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Isotype mouse IgG1 kappa
Population Doubling Time 14 hours
Name of Depositor ER Unanue
Year of Origin 1996
References

Dadaglio G, et al. Characterization and quantitation of peptide-MHC complexes produced from hen egg lysozyme using a monoclonal antibody. Immunity 6: 727-738, 1997. PubMed: 9208845

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Basic Documentation
Other Documentation
References

Dadaglio G, et al. Characterization and quantitation of peptide-MHC complexes produced from hen egg lysozyme using a monoclonal antibody. Immunity 6: 727-738, 1997. PubMed: 9208845

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm