宁波泰斯拓生物

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Eph4 1424.2

货号 TS204992
中文名称 null
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产品简介
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产品名称: Eph4 1424.2
商品货号: TS204992
Organism: Mus musculus, mouse
Tissue: breast; derived from metastatic site: lung
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 the constitutively activated form of MEK1 and can be used in MEK-MAPK pathway studies

This cell line stably expresses constitutively activated form of MEK1(MEKDD), and can be used in MEK-MAPK pathway studies.xa0

This cell line was derived from metastatic tumor and can be used to study breast cancer metastasis.xa0

Storage Conditions: liquid nitrogen vapor phase
Images: CRL-3210 Micrograph
Derivation: EpH4 1424.2 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).xa0 Cells were rederived from a metastatic tumor to the lung after selection with 1 mg/mL G418 in complete medium.
Clinical Data: female
Genes Expressed:

Glu-Glu tagged phosphorylation site MEK1 mutant (MEKDD), verified at ATCC.

constitutively activated form of MEK1, expressed

Cellular Products:


Tumorigenic: Yes
Comments:

EpH4 1424.2 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).xa0 Cells were rederived from a metastatic tumor to the lung after selection with 1 mg/mL G418 in complete medium.

This cell line stably expresses constitutively activated form of MEK1(MEKDD, Asp218/Asp222 MEK1 phosphorylation site mutant), and can be used in MEK-MAPK pathway studies. This cell line was derived from metastatic tumor and can be used to studyxa0breast cancer metastasis.xa0

Activation of MEK1 is mediated through phosphorylation of Ser218 and Ser222 by members of the Raf family of kinases.xa0

This line related to four other cell lines: B-MEKDD 116 cell line (ATCC CRL-3069), EpH4 1424 cell line (ATCC CRL-3071), EpH4 1424.1 cell line (ATCC CRL-3209), and EpH4-Ev cell line (ATCC CRL-3063).


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 and 200 mcg/mL G418
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos 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 4 X 103 to 8 X 103 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.
Subcultivation ratio: A subcultivation ratio of 1:6 to 1:15 is recommended.xa0

Medium renewal: Every 2 to 3 days

Cryopreservation: Freeze medium: complete growth medium supplemented with an additional 10% bovine calf serum and 10% DMSO
Storage temperature: liquid nitrogen vapor phase
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