PEAKrapid (ATCC® CRL-2828)

Organism: Homo sapiens, human  /  Cell Type: transformed with adenovirus 5 DNA  /  Tissue: kidney  / 

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Organism Homo sapiens, human
Tissue kidney
Cell Type transformed with adenovirus 5 DNA
Product Format frozen
Morphology epithelial
Culture Properties loosely adherent
Biosafety Level 2 [cells containing Adenovirus type 5 viral DNA sequences]
Age fetus
Applications
The cells are useful for protein expression. They support the replication of recombinant plasmids with the Epstein-Barr virus (EBV) oriP or SV40 origin of replication.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
This line was derived from the human embryonic kidney line, HEK 293, ATCC CRL-1573. 
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.05 mg/ml Gentamicin sulfate and 10% fetal bovine serum.
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. Note: The cells are loosely adherent and will detach at room temperature.
  1. Remove culture medium to a centrifuge tube.
  2. Briefly rinse the cell layer with 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 with 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. To remove trypsin-EDTA solution, transfer cell suspension to the centrifuge tube with the medium and cells from step #1 and spin at approximately 125 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
    An inoculum of 5 x 103 to 5 x 104 viable cells/mL is recommended.
  7. Place culture vessels in incubators at 37°C.
Interval: Maintain cultures at a cell concentration between 8 x 103 and 1 x 10cells/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every two to three days

Note:For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

Cryopreservation Complete growth medium supplemented with an additional 20% fetal bovine serum and 10% 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%
Note: The cells are loosely adherent and will detach at room temperature.

Population Doubling Time 26 hours
Name of Depositor J Seed
References

Nelson G, et al. An amino-acid taste receptor. Nature 416: 199-202, 2002. PubMed: 11894099

Magistrelli G, et al. Rapid, simple and high yield production of recombinant proteins in mammalian cells using a versatile episomal system. Protein Expression and Purification: 72(2): 209-216, 2010. PubMed: 20399863

Dobi A, et al. Mammalian expression cloning of nucleic acid binding proteins by agarose thin-layer gelshift clone selection. Biotechniques: 33(4): 868-872, 2002. PubMed: 12398195

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

Nelson G, et al. An amino-acid taste receptor. Nature 416: 199-202, 2002. PubMed: 11894099

Magistrelli G, et al. Rapid, simple and high yield production of recombinant proteins in mammalian cells using a versatile episomal system. Protein Expression and Purification: 72(2): 209-216, 2010. PubMed: 20399863

Dobi A, et al. Mammalian expression cloning of nucleic acid binding proteins by agarose thin-layer gelshift clone selection. Biotechniques: 33(4): 868-872, 2002. PubMed: 12398195