M1.4 (ATCC® CRL-2464)

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

Organism Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)
Cell Type hybridoma:
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
Applications
The antibody specifically binds to mouse ZP1 glycoprotein.
Storage Conditions liquid nitrogen vapor phase
Derivation
Spleen cells were fused with Sp2/0 mouse myeloma cells.
Genes Expressed
immunoglobulin; monoclonal antibody; against mouse ZP1 glycoprotein
Cellular Products
immunoglobulin; monoclonal antibody; against mouse ZP1 glycoprotein
Comments
Animals were immunized with particulate murine zonae pellucidae.
Spleen cells were fused with Sp2/0 mouse myeloma cells.
During development and maturation, mammalian oocytes are surrounded by the zona pellucida, which in the mouse is comprised of three sulfated glycoproteins, ZP1, ZP2 and ZP3.
The zona pellucida mediates sperm binding at fertilization, provides a post-fertilization block to polyspermy, and facilitates passage of pre-implantation embryos down the oviduct.
The antibody specifically binds to mouse ZP1 glycoprotein.
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 10 mM HEPES, 0.15 mg/ml oxaloacetate, 0.05 mg/ml pyruvate, 0.0082 mg/ml bovine insulin and 0.05 mM 2-mercaptoethanol, 80%; fetal bovine serum, 20%
Subculturing Cultures can be maintained by addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation of the suspension with subsequent resuspension in fresh medium. Add medium as the cell density increases. Maintain cultures at cell concentrations between 4 x 105 and 5 x 105 viable cells/mL. Attached cells may be subcultured by scraping.

Medium Renewal:  Add fresh medium every 2 to 3 days (depending on cell density)

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 IgG2a
Name of Depositor J Dean
Passage History
The zona pellucida mediates sperm binding at fertilization, provides a post-fertilization block to polyspermy, and facilitates passage of pre-implantation embryos down the oviduct.
References

Rankin TL, et al. Human ZP3 restores fertility in Zp3 null mice without affecting order-specific sperm binding. Development 125: 2415-2424, 1998. PubMed: 9609824

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Basic Documentation
References

Rankin TL, et al. Human ZP3 restores fertility in Zp3 null mice without affecting order-specific sperm binding. Development 125: 2415-2424, 1998. PubMed: 9609824

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm