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Lost of Tet-on Vector


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#1 nicoleelizabeth

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Posted 30 April 2013 - 08:01 AM

Hello, I have generated a tet-inducible model using TGFB. When I first made the cell lines it worked beautifully. Now it seems like the Tet-on vector is not regulating the expression of the TGFB. I made sure to use a cell line that had a low basal expression of TGFB and now the expression of TFGB is high in these cells before the Doxycycline is even added. Has anybody else had any problems with a Tet-on vector before? Im new to transfections and any help will be greatly appreciated!

#2 bob1

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Posted 30 April 2013 - 12:29 PM

FBS often contains tetracycline (doxycycline analogue) if it is from the EU, USA or Asia, check the source of your FBS and make sure it is tet-free. As far as I know, only New Zealand FBS is tet-free.

#3 nicoleelizabeth

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Posted 02 May 2013 - 05:51 AM

Thank you for the reply!

I am using a tet-free FBS from clontech, where I got the Tet-on Vector. I think it should be fine, it is the same FBS that I used right after transfection and it worked then, however, not it's not.

#4 bob1

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Posted 02 May 2013 - 12:22 PM

The tet-operon does also tend to be a bit leaky, though I don't think as much as you are seeing. Unfortunately I don't know of any way around this.




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