X1/5

NOTE: Both the cell line and DNA from the cell line may be available for this product. Please choose -1VL or VIAL for cells, or -DNA-5UG for DNA, 95051229, human cervix, Epithelial

• UNSPSC Code: 41106514
• Biological source: human cervix
• Relevant disease(s): cancer
• Growth mode: Adherent
• Karyotype: Not specified
• Morphology: Epithelial
• Manufacturer/tradename: ECACC

Eigenschaften

• biological source: human cervix
• manufacturer/tradename: ECACC
• packaging: tube of 5 μg 95051229-DNA-5UG, pkg of vial of cells 95051229-1VL
• growth mode: Adherent
• karyotype: Not specified
• morphology: Epithelial
• products: Not specified
• receptors: Not specified
• technique(s): cell culture | mammalian: suitable
• relevant disease(s): cancer
• shipped in: dry ice
• storage temp.: −196°C

Beschreibung

Application

Specific and stringent control of the activity of an individual gene (luciferase) in a mammalian cell can be monitored. It is also suitable for creation of

Biochem/physiol Actions

Human (HeLa) with tTA

X1/5 is a HeLa cell line stably transfected with a tet repressor fused with the activating domain of virion protein 16 of herpes simplex virus thus generating a tetracycline-controlled transactivator (tTA) This stimulates transcription from a minimal promoter sequence derived from the human cytomegalovirus promoter IE combined with tet operator sequences. Integration of a luciferase gene controlled by a tTA-dependent promoter into this tTA producing HeLa cell line leads to high levels of tetracycline-sensitive luciferase expression. The luciferase activity can be regulated over up to five orders of magnitude depending on the concentration of tetracycline in the culture medium (0-1ug/ml). Specific and stringent control of the activity of an individual gene (luciferase) in the mammalian cell can be monitored. It is also suitable for creation of on/off situations and analysis of mRNA decay rates under physiological conditions

Preparation Note

EMEM (EBSS) + 2mM Glutamine + 1% Non Essential Amino Acids (NEAA) + 1% Vitamins + 500μg/ml G418 + 300μg/ml Hygromycin + 10% Foetal Bovine Serum (FBS). Freeze cells in 40% medium, 10% Dimethyl Sulphoxide (DMSO) and 50% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:5 to 1:20 i.e. seeding at 5x1,000 to 2x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA. Alternatively cells can be washed with PBS and subcultured after addition of PBS with 0.8mM EDTA (30% of original volume)

Other Notes

Additional freight & handling charges may be applicable for Asia-Pacific shipments. Please check with your local Customer Service representative for more information.

This cell line is part of the European Collection of Authenticated Cell Cultures (ECACC), an international repository managed by the United Kingdom Health Security Agency (UKHSA). No licensing agreement is required when either this cell line or the DNA extracted from it are used for internal research purposes only. Any other use of these products is prohibited without the express written permission of UKHSA. Inquiries regarding authorized use of this cell line or its genomic DNA may be directed to culturecollections@ukhsa.gov.uk.