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CHOP-KO-DR
CHOP-KO-DR
規(guī)格:
價(jià)格:
編號(hào):B228262
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱
CHOP-KO-DR
商品貨號(hào)
B228262
Organism
Mus musculus, mouse
Tissue
embryo fibroblast/embryo
培養(yǎng)基
Cell Type
fibroblast
Product Format
frozen
Morphology
fibroblast-like
Culture Properties
adherent
Biosafety Level
2 [Cells contain 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.

Age
embryo, 13.5 days gestation
Applications
DR-Wildtype cells, CRL-2977, are available for use as a control.
These cells lack the transcription factor CHOP, a key mediator of cell death in the unfolded protein response (UPR).
CHOP is a major regulator of the endoplasmic reticulum UPR and as such, these CHOP-KO cells are useful for studying the mechanisms of UPR and integrated stress response.
Storage Conditions
liquid nitrogen vapor phase
Images
Derivation
The mouse embryonic fibroblast (MEF) cell line, CHOP-KO-DR (CRL-2979), was established from a 13.5 day-old CHOP-/- mouse embryo by SV-40 immortalization.
Genes Expressed
C/EBP homologous protein (CHOP)/growth arrest and DNA damage-inducible gene 153 (GADD153), not expressed
Cellular Products
C/EBP homologous protein (CHOP)/growth arrest and DNA damage-inducible gene 153 (GADD153), not expressed
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:
  • O.1 mM Non-Essential Amino Acids (NEAA)
  • 0.05mM 2-Mercaptoethanol
  • fetal bovine serum to a final concentration of 10%

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 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 2.0 to 3.0 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 2 X 103 to 3 X 103 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C.
Subcultivation ratio: A subcultivation ratio of 1:10 to 1:30 twice weekly is recommended.
Medium renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: fetal bovine serum, 90%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor
D Ron and H Harding
Year of Origin
1998
References

Zinszner H, et al. CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev. 12(7): 982-995, 1998. PubMed: 9531536

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