Chris H Lindley
chris at scotgate2.demon.co.uk
Wed Apr 14 16:42:03 EST 1999
On Tue, 13 Apr 1999 22:55:18 GMT, fbacha at PO-box.mcgill.ca wrote:
> I would like to inhibit translation in my TNT couple Retic. Lys. So, I ordered cycloheximide from Sigma.
>From the literature, people use it at a concentration of 10 ng/ul for RRL. My question is: in what should I resuspend
>cycloheximide and is it stable at -20??? Can I resuspend my whole 1g and keep that at -20 or should I make it fresh
>for every experiments???
I used cycloheximide to inhibit a TnT reaction. I made a stock
solution in dH2O, and added to give final conc of 5mM.
I kept it at -2odegC.
I never really used a control, so I'm not 100% sure if this
concentration really inhibited protein synthesis, but everything
turned out as I expected!!
The parameters I sed were ones which were recommended to me
as "accepted" ones to use for translation inhibition!!
Previously in a totally different set of experiments I used
cyhex at a conc of 10ug/ml (this was an in vivo experiment so
you can't really compare with the TnT kit!) But I mention it
as I stored the stock solution in aliquots at -20degC for
about 6 months and it worked exactly as it did previously!
¦ Chris H. Lindley Yorkshire, UK ¦
¦ chris at scotgate2.demon.co.uk Ferg on #os/2 and #os2uk, EFnet ¦
¦ WarpUK:UK OS/2 Users group www.warp.in-uk.net ¦
¦ Molecular Biology & OS/2 www.scotgate.demon.co.uk ¦
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