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BHK21-pcDNA3.1-HC
BHK21-pcDNA3.1-HC
規(guī)格:
貨期:
編號:B164034
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 BHK21-pcDNA3.1-HC
商品貨號 B164034
Organism Mesocricetus auratus, hamster, Syrian golden
Tissue kidney
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 cells contain CMV and SV40 DNA viral particles

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 newborn
Applications
The cells secrete recombinant human erythropoietin in media, which can be detected by ELISA assay.
This line was derived from ATCC PTA-419, which was derived from BHK-21(c-13) (ATCC CCL-10) [U.S. Pat.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
This line was derived from ATCC PTA-419, which was derived from BHK-21(c-13) (ATCC CCL-10) [U.S. Pat. 6,376,218]. The line was transfected by electroporation using the plasmid vector pcDNA3.1 containing a cDNA fragment encoding human erythropoietin. The vector contains cytomegalovirus (CMV) and SV40 viral DNA sequences and the neomycin resistance gene. The cells secrete recombinant human erythropoietin in media, which can be detected by ELISA assay.
Genes Expressed
recombinant human erythropoietin
Cellular Products
recombinant human erythropoietin
Comments
This line was derived from ATCC PTA-419, which was derived from BHK-21(c-13) (ATCC CCL-10) [U.S. Pat. 6,376,218]. The line was transfected by electroporation using the plasmid vector pcDNA3.1 containing a cDNA fragment encoding human erythropoietin. The vector contains cytomegalovirus (CMV) and SV40 viral DNA sequences and the neomycin resistance gene. The cells secrete recombinant human erythropoietin in media, which can be detected by ELISA assay.
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: 0.4 mg/ml G-418; fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; 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 0.25% (w/v) Trypsin-0.53mM 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.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:8
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor L Hsu, SC Chang
References

Hsu LW, Chang SC. Expression system for producing recombinant human erythropoietin, method for purifying secreted human erythropoietin and uses thereof. US Patent 6,376,218 dated Apr 28 2002

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

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