removal of SDS from protein samples

Phil Harrison arsphys at cc.usu.edu
Tue Jul 11 16:17:12 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.
>
>

I was using 4% SDS to solubilize some difficult proteins for 2-D electrophoresis.  Precipitating the proteins in cold acetone (it was actually 8-10 vols) removed the high concentration of SDS, which otherwise interfered with the 1st-dimension IEF gel.   It was several years ago I did this, and the only reference I had was the early (1969) Weber and Osborn paper J Biol Chem 244:4406-4412 and a conversation with a R(obert?) Poynton.  I don't remember who he was or how I found him.  Sorry I don't have more.
Further note: Maybe the NP-40 in the IEF did help tie up some residual SDS.

Phil Harrison


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