2A5 (ATCC® CRL-2444)

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

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 suspension
Biosafety Level 1
Applications
The cell line was established by Jose A. Lebron and Pamela J. Bjorkman in 1997.
Spleen cells were fused with HL-1 Friendly myeloma cells (a derivative of P3X63Ag8.653 that is deficient in both hypoxanthine phosphoribosyl transferase [HPRT] and adenine phosphoribosyl transferase [APRT]).
This antibody can be used for ELISA applications and to immunoprecipitate soluble HFE; it is not reactive in Western Blot assays.
Derivation
The cell line was established by Jose A. Lebron and Pamela J. Bjorkman in 1997.
Animals were tolerized with human beta2-microglobulin and immunized with purified, soluble, human HFE.
Spleen cells were fused with HL-1 Friendly myeloma cells (a derivative of P3X63Ag8.653 that is deficient in both hypoxanthine phosphoribosyl transferase [HPRT] and adenine phosphoribosyl transferase [APRT]).
Genes Expressed
immunoglobulin; monoclonal antibody; against HFE
Cellular Products
immunoglobulin; monoclonal antibody; against HFE
Comments
The cell line was established by Jose A. Lebron and Pamela J. Bjorkman in 1997.
Animals were tolerized with human beta2-microglobulin and immunized with purified, soluble, human HFE.
Spleen cells were fused with HL-1 Friendly myeloma cells (a derivative of P3X63Ag8.653 that is deficient in both hypoxanthine phosphoribosyl transferase [HPRT] and adenine phosphoribosyl transferase [APRT]).
The antibody recognizes the HFE portion of the HFE/beta2 heterodimer.
HFE is a major histocompatibility complex (MHC) related protein that is mutated in the iron-overload disease hereditary hemochromatosis.
This antibody can be used for ELISA applications and to immunoprecipitate soluble HFE; it is not reactive in Western Blot assays.
Complete Growth Medium HL-1 medium supplemented with 4 mM L-glutamine, 1 mM sodium pyruvate and 1% fetal bovine serum. HL-1 medium can be obtained from from Lonza (catalog number 77201).
Subculturing
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)
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 exp5 viable cells/ml.
Maintain cell density between 1 X 10 exp5 and 1 X 10 exp6 viable cells/ml.
Attached cells may be subcultured by tapping the sides of the flask until cells are dispersed.
Cryopreservation
HL-1 Complete Serum-free Medium, 75%; fetal bovine serum, 20%; DMSO, 5%.
Culture Conditions
Temperature: 37.0°C
Isotype IgM; kappa light chain
Name of Depositor PJ Bjorkman
Year of Origin 1997
References

Lebron JA, et al. Tolerization of adult mice to immunodominant proteins before monoclonal antibody production. J. Immunol. Methods 222: 59-63, 1999. PubMed: 10022372

Lebron JA, et al. Crystal structure of the hemochromatosis protein HFE and characterization of its interaction with transferrin receptor. Cell 93: 111-123, 1998. PubMed: 9546397

Basic Documentation
References

Lebron JA, et al. Tolerization of adult mice to immunodominant proteins before monoclonal antibody production. J. Immunol. Methods 222: 59-63, 1999. PubMed: 10022372

Lebron JA, et al. Crystal structure of the hemochromatosis protein HFE and characterization of its interaction with transferrin receptor. Cell 93: 111-123, 1998. PubMed: 9546397