Another option I didn't mention is presoak your centricons in 30% glycerol over 
night prior to usage.
Jürgen

On Jan 28, 2012, at 12:47 PM, Bosch, Juergen wrote:

Are you close to the theoretical isoelectric point of your protein ? Change pH 
of buffer
Jürgen

......................
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry & Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Phone: +1-410-614-4742
Lab:      +1-410-614-4894
Fax:      +1-410-955-3655
http://web.mac.com/bosch_lab/

On Jan 28, 2012, at 10:55, "rashmi panigrahi" <rashmi.panigrah...@gmail.com> 
wrote:

Hi all,
I tried to concentrate my protein using vivaspin 20  10,000 MWCO PES.
The protein was in 50mM Hepes pH 7.5, 500mM KCl  and 10% glycerol.
I lost about 90% of my protein on the membrane of the centricon.

Please  suggest some way of concentrating this protein.
Will concentrating using peg 20K be a good alternative??

regards

rashmi

......................
Jürgen Bosch
Johns Hopkins University
Bloomberg School of Public Health
Department of Biochemistry & Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Office: +1-410-614-4742
Lab:      +1-410-614-4894
Fax:      +1-410-955-2926
http://web.mac.com/bosch_lab/




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