[RASMB] triton removal

[Ines Lehner]

Dear Emma,

Triton X and other detergents with low cmc values which cannot be
removed very well by dialysis and gelfiltration can be removed using
hydrophobic polystyrene beads. There are several suppliers eg BioRads
Biobeads. For optimal absorption capacity wash them in water before use
and degass the porous beads. I use 2 times 1h for detergent removal. As
a rough guide I would use dry biobeads with 30 times the weight of the
detergent that you want to remove. 
For example: 1ml solution with 0.1% Triton, add for 1h 30mg biobeads
(shake), take off the solution and add to it another 30mg biobeads for
1h (shake).
There are several nice reviews on the subject by Rigaud in France.

Regards, Ines

On Thu, 2006-10-05 at 12:17 +0100, EJ Longman wrote:
> Does anyone have any advice on removal of triton X100 from protein samples? 
> There seem to be a number of commercially available systems specifically 
> for triton removal (as dialysis and gel filtration are not effective).
> 
> I have purified two proteins (molecular weights of  30 & 40 kDa) with 0.1% 
> triton in the buffers. The triton is not necessary for the stability of the 
> proteins but improves the purity of the protein obtained. I would like to 
> remove it as its high absorbance will cause problems in my AUC experiments.
> 
> I am also interested to know whether detergent removal can be efficient 
> enough that afterwards the sample can be used for AUC without having to 
> worry about density matching?
> 
> Thanks,
> 
> Emma
> 
> 
> ----------------------
> EJ Longman
> biejl at bristol.ac.uk
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-- 
Ines Lehner
University of Frankfurt
Institute of Biophysical Chemistry
Max von Laue Str. 9
60438 Frankfurt am Main

0049 69 798 29902