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Cell Biology
ATCC® Number: CRL-2444™    Price: $399.00
Designations: 2A5
Depositors:  PJ Bjorkman
Isotype: IgM; kappa light chain
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: suspension
Organism: Mus musculus (B cell); Mus musculus (myeloma) (mouse (B cell); mouse (myeloma))
Morphology: lymphoblast
Source: Cell Type: hybridoma: B lymphocyte;
Cellular Products: immunoglobulin; monoclonal antibody; against HFE
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
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.
Propagation: ATCC 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 BioWhittaker, Walkersville, MD 21793, 800-638-7184.
Temperature: 37.0°C
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.
Preservation: HL-1 Complete Serum-free Medium, 75%; fetal bovine serum, 20%; DMSO, 5%.
Related Products: recommended serum:ATCC 30-2020
References: 39791: Lebron JA, et al. Tolerization of adult mice to immunodominant proteins before monoclonal antibody production. J. Immunol. Methods 222: 59-63, 1999. PubMed: 10022372
39792: 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
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