HP6058 (ATCC® CRL-1786)

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

Permits and Restrictions

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Organism Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
Cell Type hybridoma: B lymphocyte
Product Format frozen
Morphology lymphoblast
Culture Properties mixed, adherent and suspension
Biosafety Level 1
Applications
Tested and found negative for ectromelia virus (mousepox).
Derivation
Animals were immunized with purified human myeloma IgG.
Spleen cells were fused with Sp2/0-Ag14 myeloma cells.
Genes Expressed
immunoglobulin; monoclonal antibody; against human IgG (FC) of subclasses 1, 2 and 3 (IgG1, IgG2, IgG3)
Cellular Products
immunoglobulin; monoclonal antibody; against human IgG (FC) of subclasses 1, 2 and 3 (IgG1, IgG2, IgG3)
Comments
Animals were immunized with purified human myeloma IgG.
Spleen cells were fused with Sp2/0-Ag14 myeloma cells.
Tested and found negative for ectromelia virus (mousepox).
Complete Growth Medium Dulbecco's modified Eagle's medium with 4.5 g/L glucose, 10 mM HEPES and 0.05 mM 2-mercaptoethanol, 90%; fetal bovine serum, 10%
Subculturing
Protocol: Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 1. Remove culture medium with floating cells to a centrifuge tube. 2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor. 3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 10 minutes). 4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. 5. To remove trypsin-EDTA solution, transfer cell suspension to the centrifuge tube with the medium and cells from step #1 and spin at approximately 125 xg for 5 to10 minutes. 6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Resuspend cells at 2 X 10(5) viable cells/ml 7. Place culture vessels in incubators at 37°C.
Medium Renewal: Every 2 to 3 days
Culture Conditions
Temperature: 37.0°C
Isotype IgG1
Name of Depositor CB Reimer
References

Reimer CB, et al. Evaluation of thirty-one mouse monoclonal antibodies to human IgG epitopes. Hybridoma 3: 263-275, 1984. PubMed: 6209201

Jefferis R, et al. Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: Results of an IUIS/WHO collaborative study. Immunol. Lett. 10: 223-252, 1985. PubMed: 3899923

Basic Documentation
Restrictions

The depositor has provided these cells for research purposes only and with the following restrictions: (1) no distribution should be made to third parties, (2) any proposed commercial use must be negotiated with the Centers for Disease Control, (3) all papers reporting any use of these or derived clones should make direct reference to the original publication (Hybridoma 3:263-275, 1984).

References

Reimer CB, et al. Evaluation of thirty-one mouse monoclonal antibodies to human IgG epitopes. Hybridoma 3: 263-275, 1984. PubMed: 6209201

Jefferis R, et al. Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: Results of an IUIS/WHO collaborative study. Immunol. Lett. 10: 223-252, 1985. PubMed: 3899923