M1/9.3.4.HL.2

Code: 0153
Cell Line: M1/9.3.4.HL.2
Species: Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)

Vulgar Name: Rat/mouse
Tissue: Spleen
Morphology: Lymphoblast
Growth Properties: Suspension
Derivation: Rats were immunized with C57BL/10 mouse spleen cells enriched for T lymphocytes. Spleen cells were fused with NS-1 myeloma cells. The antibody reacts with an antigen, CD45, present on mouse lymph node and spleen cells but not on brain, kidney, liver or red blood cells. It antibody reacts with all isoforms of CD45. The CD45 antigen is common to all lines of leukocyte differentiation.
Applications: The antibody reacts with an antigen, CD45, present on mouse lymph node and spleen cells but not on brain, kidney, liver or red blood cells. It antibody reacts with all isoforms of CD45. The CD45 antigen is common to all lines of leukocyte differentiation.
Products: "immunoglobulin; monoclonal antibody; against mouse common leukocyte antigen (200000 dalton, CD45). Genes Expressed: immunoglobulin; monoclonal antibody; against mouse common leukocyte antigen (200000 dalton, CD45)"
Biosafey: 1
Culture Medium:
Complete growth medium: The base medium for this cell line is Dulbecco's Modified Eagle's Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

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.


Medium Renewal: Every 2 to 3 days

Subcultivation ratio: Maintain cell density between 1 X 10(5) and 1 X 10(6) viable 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:
Springer T, et al. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur. J. Immunol. 8: 539-551, 1978. PubMed: 81133

Stern PL, et al. Monoclonal antibodies as probes for differentiation and tumor-associated antigens: a Forssman specificity on teratocarcinoma stem cells. Cell 14: 775-783, 1978. PubMed: 567532

Springer TA. Monoclonal antibody analysis of complex biological systems. Combination of cell hybridization and immunoadsorbents in a novel cascade procedure and its application to the macrophage cell surface. J. Biol. Chem. 256: 3833-3839, 1981. PubMed: 7217058

Luqman M, et al. Differential expression of the alternatively spliced exons of murine CD45 in Th1 and Th2 cell clones. Eur. J. Immunol. 21: 17-22, 1991. PubMed: 1671357

Springer TACell -surface differentiation in the mouse: Characterization of Jumping and Lineage antigens using xenogeneic rat monoclonal antibodiesIn: Springer TAMonoclonal AntibodiesNew YorkPlenum Presspp. 185-217, 1980

Milstein C, et al. Monoclonal antibodies and cell surface antigens. Cell Biol. Int. Rep. 3: 1-16, 1979. PubMed: 88272
Depositors: Banco de Células do Rio de Janeiro
ATCC: TIB-122