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HP6058
HP6058
規(guī)格:
貨期:
編號:B164643
品牌:Mingzhoubio

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產(chǎn)品名稱 HP6058
商品貨號 B164643
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

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Applications
Tested and found negative for ectromelia virus (mousepox).
Storage Conditions liquid nitrogen vapor phase
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 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 x g for 5 to 10 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 105 viable cells/m.
  7. Place culture vessels in incubators at 37°C.
Medium Renewal Every 2 to 3 days
Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Isotype IgG1
Name of Depositor CB Reimer
Deposited As mouse (B cell); mouse (myeloma)
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

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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