M17/4.4.11.9 (new clone of M17/4.2) (ATCC® TIB-217)

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
Monoclonal antibodies against LFA-1 can block killing by inhibiting the adhesion between cytolytic T cells and target cells.
The LFA-1 and Mac-1 antigens are composed of two chains or subunits, alpha and beta.
Storage Conditions liquid nitrogen vapor phase
Derivation
Spleen cells were fused with NS-1 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 alpha subunit of LFA-1 (mouse cytotoxic T cell antigen, CD11a)
Cellular Products
immunoglobulin; monoclonal antibody; against the alpha subunit of LFA-1 (mouse cytotoxic T cell antigen, CD11a)
Comments
Animals were immunized with cytotoxic T cells from C57BL/6 mice immunized with P815 cells.
Spleen cells were fused with NS-1 myeloma cells.
LFA-1 is a cell surface antigen found on cytolytic T lymphocytes.
Monoclonal antibodies against LFA-1 can block killing by inhibiting the adhesion between cytolytic T cells and target cells.
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.
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

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 5 X 10(4) viable cells/ml.
Interval: Maintain between 5 X 10(4) and 5 X 10(5) cells/ml.
Medium Renewal: Every 3 to 4 days
Cryopreservation
Freeze medium: Complete growth medium supplemented with 7.5% (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

Sanchez-Madrid F, et al. Antigens involved in mouse cytolytic T-lymphocyte (CTL)-mediated killing: functional screening and topographic relationship. Cell. Immunol. 73: 1-11, 1982. PubMed: 6983917

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

Sanchez-Madrid F, et al. Antigens involved in mouse cytolytic T-lymphocyte (CTL)-mediated killing: functional screening and topographic relationship. Cell. Immunol. 73: 1-11, 1982. PubMed: 6983917