Removal of SDS from protein for structural studies

Tom Branham tbranham at mm.wellesley.edu
Mon May 22 09:39:30 EST 1995


Hiranya Roychowdhury (hroychow at NMSU.EDU) wrote:
> On 18 May 1995, Dr. Jeff Lightfoot wrote:

> > What methods are recommended for removing 1% SDS from protein?   Is
> > dialyzing sufficient?  The protein is to be used for crystallography and/or
> > nmr.  Pierce sells a gel that is supposed to remove detergents, but they
> > keep bumping the availability date back.  Thanks for your help.
> > 
> > Jeff.


> Removal of SDS from protein can be accomplished in a number of different 
> ways:
> 	1. Electrodialysis using a dialysis bag. Ofcourse the surrounding 
> buffer has to be free of diaysis.
> 	2. Equilibrium dialysis, which you have mentioned.
> 	3. Electroelution into SDS free buffer using Triton X100, where 
> TritonX100 is removed by dialysis later. (Holloway,('73) Anal. Biochem., 
> 53:304)
> 	4. Ion retardation columns (Kapp & Vinogradov (78), Anal. 
> Biochem. 91: 230.
> 	5. Anion Exchange, Weber & Cutter JBC 246:4504.


If you want to use the protien for nmr or xray afterwards, can we assume
you are still expecting it to be folded??????????
You might be better off trying that from a native gel rather than SDS,
or use some other technique for separation (Perhaps isoelectic focusing).


--
tom
tbranham at mm.wellesley.edu




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