| 產品名稱 |
EpH4 1424.1 |
| 商品貨號 |
B230243 |
| Organism |
Mus musculus, mouse |
| Tissue |
breast epithelium (mammary gland);
derived from metastatic site:
kidney |
| Cell Type |
epithelial cell |
| Product Format |
frozen |
| Morphology |
epithelial-like |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain CMV and SV40 viral DNA sequences]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
breast cancer |
| Gender |
female |
| Applications |
This cell line stably expresses constitutively activated form of MEK1 (Asp218/Asp222 MEK1 phosphorylation site mutant) and can be used in MEK-MAPK pathway studies.
This cell line is tumorigenic, and can be used to study a breast cancer in vivo model
This cell line was derived from metastatic tumor and can be used to study breast cancer metastasis. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
EpH4 1424.1 was derived from BALB/c mouse xenografts initiated with EpH4 cells stably transfected with an expression vector containing Glu-Glu epitope-tagged phosphorylation site MEK1 mutant (MEKDD). Cells were rederived from a metastatic tumor to the kidney after selection with 1 mg/mL G418 in complete medium. |
| Genes Expressed |
Constitutively activated form of MEK1, expressed |
| Cellular Products |
stably expresses Glu-Glu tagged MEKDD, which is a constitutively activated form of MEK1 (Asp218/Asp222 MEK1 phosphorylation site mutant) |
| Tumorigenic |
Yes |
| Comments |
EpH4 1424.1 was derived from BALB/c mouse xenografts initiated with EpH4 cells stably transfected with an expression vector containing Glu-Glu epitope-tagged phosphorylation site MEK1 mutant (MEKDD). Cells were rederived from a metastatic tumor to the kidney after selection with 1 mg/mL G418 in complete medium.
Activation of MEK1 is mediated through phosphorylation of Ser218 and Ser222 by members of the Raf family of kinases.
This cell line stably expresses constitutively activated form of MEK1(MEKDD), and can be used in MEK-MAPK pathway studies.
This cell line was derived from metastatic tumor and can be used to study breast cancer metastasis.
There are four additional related cell lines: B-MEKDD 116 cell line (ATCC CRL-3069), EpH4 1424 cell line (ATCC CRL-3071), EpH4 1424.2 cell line (ATCC CRL-3210) and EpH4-Ev cell line (ATCC CRL-3063).
The EpH4 1424.1 cell line produces the constitutively activated MEK1 mutant MEKDD which has been tagged with Glu-Glu, verified at ATCC. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated DMEM Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
- 10% Bovine Calf Serum
- 1.2 mcg/mL Puromycin
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 x 103 to 8 x 103 viable cells/cm2 is recommended.
- Incubate cultures at 37°C. Subculture when the cell concentration is between 8 x 104 to 1.5 x 105 cells/cm2.
Subcultivation ratio: A subcultivation ratio of 1:6 to 1:15 is recommended.
Medium renewal: Every 2 to 3 days |
| Culture Conditions |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Name of Depositor |
P Leder |
| References |
Pinkas J, et al. MEK1 signaling mediates transformation and metastasis of EpH4 mammary epithelial cells independent of an epithelial to mesenchymal transition. Cancer Res. 62(16): 4781-90. PubMed: 12183438
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