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 <18.104.22.168.20000710091753.0081b100 at cc.usu.edu>, Phil Harrison
<arsphys at cc.usu.edu> wrote:
> At 11:40 AM 07/10/2000 +0000, Mark Bond wrote:
> >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
> 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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