removal of SDS from protein samples

john f hess jfhess at ucdavis.edu
Tue Jul 11 14:30:00 EST 2000


Acetone ppt'n removes SDS?  I wouldn't expect that.  I would expect
that the protein and the SDS would ppt.  Got a handy Reference?

I have used TX-100 to remove SDS from proteins.  Of course, it helps if
you don't go over board with the SDS, but an excess of TX100 will pull
the SDS off.  A reference?  Don't have one, unfortunately.  1% TX might
be too much for some assays, but depending on the assay, might be OK. 
All you can do is try it.




In article <3.0.6.32.20000710091753.0081b100 at cc.usu.edu>, Phil Harrison
<arsphys at cc.usu.edu> wrote:

> At 11:40 AM 07/10/2000 +0000, Mark Bond wrote:
>  
> >Hi,
> >
> >Does anyone know of an easy way to removal SDS from protein samples. I know
> >you can get detergent out columns from genotech that remove free sds but how
> >can i remove SDS bound to the protein?  Can I add triton - x- 100 ? This
> >should form micells and pull the SDS from the protein into the hydrophobic
> >micells, shouldn't it? Can I do enzyme assays with the triton there?
> >
> 
> A better solution would be to add 4 volumes of cold acetone and precipitate
> the protein.
> Add 5% sucrose to keep the protein from becoming completely dehydrated and
> insoluble.
> The SDS remains dissolved in the acetone and is removed from the protein. 
> That seems
> to me to be a better solution than adding another substance that may
> interfere with your
> enzymes (the SDS almost surely will, the Triton-X, very possibly). 
> 
> Phil Harrison
> 
> 
> ---






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