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Sigma

X1/5

95051229

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Properties

Related Categories Cell Culture, ECACC Cell Lines, S-Z More...
biological source   human cervix
growth mode   Adherent
karyotype   Not specified
morphology   Epithelial
products   Not specified
receptors   Not specified
application(s)   cell culture | mammalian: suitable
relevant disease(s)   cancer
shipped in   dry ice
storage temp.   −196°C

Description

Cell Line Origin

Human (HeLa) with tTA

Cell Line Description

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

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

Culture Medium

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).

Subculture Routine

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

Cultures from HPA Culture Collections and supplied by Sigma are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.

Depositor and originator: Dr M Gossen and Prof Dr H Bujard, Zentrum für Moleculaire Biologie, University Heidelberg

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

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Cell Culture Guide

ECACC Laboratory Handbook
Protocols & Articles
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