GuCl in SDS Page

D.Lippert lippy at UVic.CA
Thu Nov 21 11:53:23 EST 1996


bijgh at zeus.bris.ac.uk (Jared Head) wrote:

>Hi,

>Can anybody tell me a quick way of prepping samples of protein in 
>guanadinium chloride, so that they will run on SDS PAGE.  The GuCl seems 
>to precepitate when mixed with SDS.  I've only got small samples so dialysis 
>will be difficult.

>Cheers,

>Jared

>-- 
>Jared Head     at the Department of Biochemistry, University of Bristol

>  "Happily unaware of the disadvantages of being a terrible writer with 
>                              nothing to say..."

I would suggest using 8Murea instead.  It will work just as well as
GuCl and does not precipitate SDS.  You don't even have to remove it
from your sample.  Since the urea is uncharged, it will sit in the
sample well and doesn't interfere with the running of the gel.  I've
used this technique in the past with no problems.  However, if you
wish to do sequencing on this material, be aware that urea can block
the N-terminus.  To avoid this, acidify the urea prior to using it (pH
6.0 is acidic enough). This destroys the urea breakdown product that
will block your N-terminal.  Hope you have better luck.

Dustin




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