question about cycloheximide

Nico Dantuma nico.dantuma at mtc.ki.se
Wed Aug 26 09:03:00 EST 1998


Hi,

I tried in an experiment to inhibit protein synthesis with cycloheximide.
To my surprise I didn't see any effect of the cycloheximide incubation.
However I am not completely sure abou this since it could also be due to my
monitoring system (GFP fusion in FACS analysis). Since I have never used
cycloheximide I did a simple experiment to test the activity of
cycloheximide by incubating a confluent HeLa culture overnight with 100
ug/ml cycloheximide. I expected a high mortality because of the
cycloheximide but I didn't observe a clear difference with a control culture.

1) Can I conclude from the latter experiment that my cycloheximide is not
inhibiting protein synthesis under the conditions that I used?

2) If so, what did I do wrong? I made a 10 mg/ml stock solution in PBS,
filter sterilized it and diluted it 100 fold in the incubation medium. Do I
have to add a supplement?

Thanks in advance,

Nico Dantuma
------------------------------------------
Nico Dantuma
Microbiology and Tumor Biology Center
Karolinska Institute
Box 280
S-17177 Stockholm
Sweden
Phone: + 46 8 7286281
Fax: +46 8 331399 	



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