cycloheximide???

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:
>Dear Readers,
>
>	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!

Cheers
Chris

-- 
ATGCTGCTAGTCGTAGCATGCTGCTTGATCGATGCGGTACGTGATGATCGTAGCTAGCTGGGCTAGTGG
¦  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  ¦
TACGACGATCAGCATCGTACGACGAACTAGCTACGCCATGCACTACTAGCATCGATCGACCCGATCACC




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