127H细胞
数量: 大量
生长状态: 悬浮生长
是否是肿瘤细胞: 0
物种来源: 小鼠
运输方式: 冻存运输
器官来源: 脾
ATCC Number: CRL-2738™
细胞形态: **样
**类型: IgG1 and 2b kappa
127H细胞Designations: 133-10F6
Depositors: National Cancer Institute
Isotype: IgG1 and 2b kappa
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: suspension
Organism: Mus musculus (B cell); Mus musculus (myeloma) deposited as mouse (B cell); mouse (myeloma)
Morphology: lymphoblast
Source: Organ: spleen
Cell Type: 127H细胞hybridoma: B lymphocyte;
Cellular Products: immunoglobulin; monoclonal antibody; against a synthetic c-myb oncogene peptide
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Tumorigenic: Yes
Comments: Animals were immunized with a synthetic polypeptide with the sequence LGEHHCTPSPPVDHG corresponding to peptides 348 to 363 of the c-myb oncogene, formerly v-myb (residues 160 to 175). Spleen cells were fused with Sp2/0 myeloma cells. Using immunoblot techniques, the ascites produced from these cells recognized proteins of approximately 45,000 and 75,000 daltons in a variety of transformed mammalian cells. In addition, proteins of 45,000 and 55,000 were detected in various urine samples. The IgG1 and IgG2b reactivities of the antibody with the c-myb peptide (348 to 363) were confirmed by ELISA. 127H细胞IgG1 was strongly positive while IgG2b was only slightly reactive. A culture submitted to the ATCC in February of 2001 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol: 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(5) viable cells/ml.
Interval: Maintain cell density between 1 X 10(5) and 1 X 10(6) viable cells/ml.
Medium Renewal: 127H细胞Add fresh medium every 2 to 3 days (depending on cell density)
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase