127H細胞

數量： 大量

生長狀態： 懸浮生長

是否是腫瘤細胞： 0

物種來源： 小鼠

運輸方式： 凍存運輸

器官來源： 脾

ATCC Number： CRL-2738?

細胞形態： **樣

**類型： IgG1 and 2b kappa

127H細胞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細胞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細胞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細胞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