人妻少妇偷人精品无码丨色婷婷av久久久久久久丨欧美xxxx做受性欧美88丨欧洲女人牲交视频免费丨亚洲精品久久久av无码专区

B-MEKDD 116 was produced by transfecting parental EpH4, a mouse mammary epithelial cell line, with an expression vector containing Glu-Glu epitope-tagged phosphorylation site MEK1 mutant (MEKDD).  After selection in complete medium supplemented with 1 mg/mL G418 for 14 days, individual clones were isolated and expanded.

B-MEKDD 116 was produced by transfecting parental EpH4, a mouse mammary epithelial cell line, with an expression vector containing Glu-Glu epitope-tagged phosphorylation site MEK1 mutant (MEKDD).  After selection in complete medium supplemented with 1 mg/mL G418 for 14 days, individual clones were isolated and expanded.

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 build a breast cancer in vivo model. There are four additional related cell lines: EpH 1424 cell line (ATCC CRL-3071), EpH4 1424.1 cell line (ATCC CRL-3209), EpH4 1424.2 cell line (ATCC CRL-3210), and EpH4-Ev cell line (ATCC CRL-3063).

The B-MEKDD 116 cell line produces the constitutively activated MEK1 mutant MEKDD which has been tagged with Glu-Glu, verified at ATCC.

1. Remove and discard culture medium.
2. 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.
3. 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.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 8 x 10³ to 1.5 x 10⁴ viable cells/cm² is recommended.
6. Incubate cultures at 37°C. Subculture when the cell concentration is between 8 x 10⁴ to 1.5 x 10⁵ cells/cm².

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

Pinkas J, et al. Bcl-2-mediated cell survival promotes metastasis of EpH4 betaMEKDD mammary epithelial cells. Mol. Cancer Res. 2(10): 551-6, 2004. PubMed: 15498929