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283TAg

規格:

價格:

編號:B206505

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

規格:

凍干粉

斜面

甘油

平板

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產品名稱

283TAg

商品貨號

B206505

Organism

Mus musculus, mouse

Tissue

embryo

培養基

Cell Type

fibroblast immortalized with SV40 large T antigenSV40 large T antigen transfected

Product Format

frozen

Morphology

fibroblast

Culture Properties

adherent

Biosafety Level

2 cells containing 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

14.5 day gestation embryo

Applications

DNA repair studies

Storage Conditions

liquid nitrogen vapor phase

Images

Derivation

283TAg (Polb/Mpg double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/ N-methylpurine-DNA glycosylase (Mpg, Aag) double null embryos at day 14.5 of gestation. The cell line was established by transfection with an expression vector for SV40 large T antigen [PubMed: 8538772]. The cells are transgenic for lambda LIZ (Lac I/cII).

Comments

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%.

Subculturing

Protocol:

Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
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.
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 10(3) to 4 X 10(4) viable cells/cm2 is recommended.
Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 6 X 10(3) and 1 X 10(5) cells/cm2.

Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended

Medium Renewal: Two to three times weekly

Cryopreservation

Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Population Doubling Time

29 hours

Name of Depositor

RW Sobol

Year of Origin

2000

References

Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772

Sobol RW, et al. Base excision repair intermediates induce p53-independent cytotoxic and genotoxic responses. J. Biol. Chem. 278: 39951-39959, 2003. PubMed: 12882965

Engelward BP, et al. Base excision repair deficient mice lacking the Aag alkyladenine DNA glycosylase. Proc. Natl. Acad. Sci. USA 94: 13087-13092, 1997. PubMed: 9371804

Sobol RW, et al. Mutations associated with base excision repair deficiency and methylation-induced genotoxic stress. Proc. Natl. Acad. Sci. USA 99: 6860-6865, 2002. PubMed: 11983862

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