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0002 - 145-2C11

BCRJ Code 0002
Cell Line 145-2C11
Species Cricetulus migratorius (B cell); Mus musculus (myeloma), hamster, Armenian (B cell); mouse (myeloma)
Vulgar Name Hamster / Mouse
Tissue Blood
Cell Type Hybridoma: B Lymphocyte
Morphology Lymphoblast-Like
Growth Properties Suspension
Derivation Animals were immunized with the BM10-37 mouse cytotoxic T lymphocyte (CTL) cell clone (anti H-2 Kb). Spleen cells were fused with Sp2/0-Ag14 myeloma cells.
Applications It reacts with all mature T cells and can both activate and inhibit T cell function. The antibody is specific for a 25000 dalton protein component (CD3 epsilon) of the antigen specific T cell receptor. The antibody reacts with the murine T cell receptor (CD3 - T3) complex. The antibody does not react with peripheral blood lymphocytes from rats, rabbits, miniature swine or hamsters. It reacts with all mature T cells and can both activate and inhibit T cell function.
Products immunoglobulin; monoclonal antibody; against mouse CD3
Biosafety 1
Addtional Info The origin of this cell line should be acknowledged in all relevant publications. May be distributed to scientific institutions; not to be distributed for any comm
Culture Medium DMEM with 4 mM L-glutamine, 4500 mg/L glucose and 10% of fetal bovine serum.
Subculturing Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10e5 viable cells/mL. Maintain cultures at a cell concentration between 1 x 10e5 and 1 x 10e6 cells/mL. NOTE: Do not allow the cell concentration to exceed 1 x 10e6 cells/mL.
Subculturing Medium Renewal Every 2 to 3 days
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
References Leo O, et al. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc. Natl. Acad. Sci. USA 84: 1374-1378, 1987. PubMed: 2950524 Kayagaki N, et al. Polymorphism of murine Fas ligand that affects the biological activity. Proc. Natl. Acad. Sci. USA 94: 3914-3919, 1997. PubMed: 9108079
Depositors Alberto Nobrega, Universidade Federal do Rio de Janeiro