immunoglobulin; monoclonal antibody; against E. coli O157 O-antigen
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Tumorigenic:
Yes
Comments:
Animals were immunized with whole bacterium antigen from E. coli O157:H7 (ATCC 43895). Spleen cells were fused with P3X63Ag8.653 mouse myeloma cells. The antibody is specific for E. coli O157 O-antigen and for the Salmonella serogroup N O-antigen (which is identical to the E. coli O157 antigen). Both ascites and culture media react with extremely high affinity. High titer ascites can be produced but these cells become lethal to mice within approximately 5 days post-intraperitioneal injection. The antibody has been used in ELISA, Western Blot, lateral flow and chemiluminescense assays.
ATCC complete growth medium: Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.1 mM nonessential amino acids, 80%; heat-inactivated fetal bovine serum, 20% Temperature: 37.0°C
Subculturing:
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density) 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 X 10 exp5 viable cells/ml. Maintain cell density between 1 X 10 exp5 and 1 X 10 exp6 viable cells/ml.
Preservation:
culture medium 95%; DMSO, 5%
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
References:
46718: Westerman RB, et al. Production and characterization of monoclonal antibodies specific for the lipopolysaccharide of Escherichia coli O157. J. Clin. Microbiol. 35: 679-684, 1997. PubMed: 9041412