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Code: 0208
Cell Line: R24
Species: Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
Morphology: Lymphoblast
Growth Properties: Suspension
Derivation: Spleen cells were fused with NS-1 myeloma cells.
Applications: The antibody reacts with the M-18 antigen system
Products: immunoglobulin; monoclonal antibody; against a human melanoma cell line (M-18 antigen system)
Biosafety: 1
Additional info: Animals were immunized with the SK-MEL-28 human melanoma cell line. Spleen cells were fused with NS-1 myeloma cells. The antibody reacts with the M-18 antigen system.
Culture Medium: RPMI 1640 with 2.0 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10.0 mM HEPES, and 1.0 mM sodium pyruvate, 90%; fetal bovine serum, 10%.
Subculturing: Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 2 X 10 exp5 cells/ml and maintain between 1 X 10 exp5 and 1 X 10 exp6 cells/ml.
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: J.Exp.Med. 155:1133-1147, 1982 ibid., 156:1755-1766, 1982 Behring Inst. Mitt. 74:14-18, 1984 Cancer REs. 190-194, 1984 Eur. J. Clin. Onc. 21:907-912, 1985 Proc. Natl. Acad. Sci. USA 82:1242-1246, 1985
Depositors: Banco De Células Do Rio De Janeiro
ATCC: HB-8445