ultrafiltration to v. small volume

lgb at ibcp.fr lgb at ibcp.fr
Sat Nov 11 07:22:12 EST 1995


In article <47ufkq$5ao at net.auckland.ac.nz>, surus at ccu1.auckland.ac.nz
(Surus) wrote:

> 
> I am trying to find the most efficient way of concentrating a small volume of
> membrane protein solution containing detergent to a volume of c. 100-200 ul.
> As the protein of interest exists in small concentration, the device used must
> have v. low protein absorption characteristics. I cannot increase the conc. of
> detergent ( this excludes speedvac or lyophilisation?).
> Does anyone know whether there are available ultrafiltration devices (10,000-
> 30,000 MW cutoff) for such small volumes? Or perhaps there is some other way
> of solving this problem. Many thanks, Andre.

I suggest you to try the ultrafiltration series sold by Amicon. You can
concentrate your sample by Centriprep (from 15 ml to 0.5 ml), Centricon (2
ml to  25-50 ul) or Microcon (0.5 ml to 10 ul). You can concentrate your
sample and completely recover your protein. I routinely use these devices
to concentrate both soluble and memebrane proteins. But be very careful to
choose the MWCO, esp. when you work on membrane proteins. The detergent
micelle should be taken into account when you choose the membrane MWCO. 

Good luck.

M. Dong
lgb at ibcp.fr



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