, September 03, 2009 5:05 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Devices Suitable to Concentrate Protein Solutions
in Liter Scale
Dear Dr Ku,
It was a wonderful idea! However, the refolding buffer contained 500mM
L-Arg and 1mM EDTA.
And I want to try other affinity columns
[mailto:ccp...@jiscmail.ac.uk] On Behalf Of Xuan
Yang
Sent: Thursday, September 03, 2009 5:05 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Devices Suitable to Concentrate Protein Solutions in
Liter Scale
Dear Dr Ku,
It was a wonderful idea! However, the refolding buffer contained
may be diafiltration devices might work for you. the membranes are
kind of expensive but we had used this effectively to reduce the volume
of cell culture media where we had secreted proteins in large volumes
of culture media.
here is a link
http://www.spectrapor.com/lit/hfdial.pdf
padayatti
On
: radovan.spu...@med.kuleuven.be
From: CCP4 bulletin board [ccp...@jiscmail.ac.uk] On Behalf Of Yingyun Liu
[yingyun...@jhu.edu]
Sent: Thursday, September 03, 2009 16:48
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Devices Suitable to Concentrate Protein
From: CCP4 bulletin board [ccp...@jiscmail.ac.uk] On Behalf Of Yingyun Liu
[yingyun...@jhu.edu]
Sent: Thursday, September 03, 2009 16:48
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Devices Suitable to Concentrate Protein Solutions in
Liter Scale
An anion exchange column with a large
Can you put a 6His-tag on your protein and add some Ni-NTA beads to
your solution to capture (and concentrate) the properly folded
molecules?
Quoting Xuan Yang pattisy...@gmail.com:
Dear All,
I am working on protein refolding via dialysis in large volumn
(typically 2~4 litters). It was
Dear Dr Ku,
It was a wonderful idea! However, the refolding buffer contained 500mM L-Arg
and 1mM EDTA.
And I want to try other affinity columns, just don't know what resin would
be appropriate.
Sincerely,
Xuan Yang
2009/9/3 Shao-Yang Ku s...@embl-hamburg.de
Can you put a 6His-tag on your
Dear Xuan Yang,
for such large volumes we use a Pellicon type system. The one we have in
the lab is a polyethersulfone PTJK prep/Scale TFF 2.5 FT2 High flux 10 k
from Millipore.
good luck
Demetres
Xuan Yang wrote:
Dear All,
I am working on protein refolding via dialysis in large volumn
An anion exchange column with a large enough bed volume might be used to
concentrate the protein. You probably need to change the pH of the
solution so that the protein sticks to the column.
On Thu, 2009-09-03 at 17:05 +0800, Xuan Yang wrote:
Dear Dr Ku,
It was a wonderful idea! However,
1. Yes, Ion Exchange (not just Anion as we don't know anything about
their protein (unless I missed an email)) is a great capture step if you
have the time to mess about.
2. When I have been refolding recently, after the refolding step we have
found that a good, old-fashioned Ammonium sulphate
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