M18/2.a.12.7 (new clone of M18/2.a.8) (ATCC® TIB-218)

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

Permits and Restrictions

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Organism Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)
Cell Type hybridoma: B lymphocyte
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
Applications
The LFA-1 antigen is associated with antigen specific T lymphocyte mediated cell killing.
The LFA-1 and Mac-1 antigens are composed of two chains or subunits, alpha and beta.
Tested and found negative for ectromelia virus (mousepox).
Storage Conditions liquid nitrogen vapor phase
Derivation
Spleen cells were fused with P3X63Ag8.653 myeloma cells.
The line was recloned by TA Springer in 1988, and a new seed stock was created at the ATCC.
Genes Expressed
immunoglobulin; monoclonal antibody; against the beta subunit of LFA-1 and Mac-1 (CD18)
Cellular Products
immunoglobulin; monoclonal antibody; against the beta subunit of LFA-1 and Mac-1 (CD18)
Comments
Animals were immunized with mouse cytotoxic T cell glycoproteins.
Spleen cells were fused with P3X63Ag8.653 myeloma cells.
The Mac-1 antigen is a macrophage differentiation antigen associated with type three complement receptor (CR3).
The LFA-1 antigen is associated with antigen specific T lymphocyte mediated cell killing.
The LFA-1 and Mac-1 antigens are composed of two chains or subunits, alpha and beta.
The beta chain of LFA-1 (Leukocyte Function Associated antigen) is immunologically identical to the beta chain of the Mac-1 antigen.
Tested and found negative for ectromelia virus (mousepox).
The line was recloned by TA Springer in 1988, and a new seed stock was created at the ATCC.
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium:
  • fetal bovine serum to a final concentration of 10%
  • 0.05 mM 2-mercaptoethanol

The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium:
  • fetal bovine serum to a final concentration of 10%
  • 0.05 mM 2-mercaptoethanol

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: Every 2 to 3 days
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 rat IgG2a kappa
Name of Depositor TA Springer
References

Sanchez-Madrid F, et al. Mapping of antigenic and functional epitopes on the alpha-and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and MAC-1. J. Exp. Med. 158: 586-602, 1983. PubMed: 6193226

Basic Documentation
Restrictions

Please acknowledge the origin of this cell line in all relevant publications by citing the following publication(s):

References

Sanchez-Madrid F, et al. Mapping of antigenic and functional epitopes on the alpha-and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and MAC-1. J. Exp. Med. 158: 586-602, 1983. PubMed: 6193226