Hi Nazia,
Answer to your last email about handling the detergents. It is suggested
for all the detergent powders to be stored at -20 in a desiccator box or
simply the bottle should be properly sealed with parafilm since the
detergent powders are very hygroscopic and quickly absorb moisture. There
Dear all,
Thank you for the prompt and valuable suggestions. I had made a typo in my
earlier mail. I use 50mM NaCl, not 5mM NaCl. I am facing this problem only
while using DDM in buffers. My other runs, even with 8M urea run perfectly
fine.
I will try increasing the salt concentartion and degas b
If your buffer can't go through a 0.2 micron filter easily, you
shouldn't run it through your FPLC. Detergent purity may be an issue.
I also experienced problems filtering a buffer containing CHAPS from
vendor X. When I switched to Anatrace CHAPS, no more clogging.
Ho
Ho Leung Ng
University
f Nazia
> Nasir Phd2009,ProteinCrystall.Lab [nazia.nasi...@nii.ac.in]
> *Sent:* Friday, July 26, 2013 1:45 PM
> *To:* CCP4BB@JISCMAIL.AC.UK
> *Subject:* [ccp4bb] Off topic: Gel filtration of membrane protein
>
> We are trying to purify a membrane protein using different detergents
> (D
ystall.Lab [nazia.nasi...@nii.ac.in]
Sent: Friday, July 26, 2013 1:45 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Off topic: Gel filtration of membrane protein
We are trying to purify a membrane protein using different detergents (DDM, OG
etc.). We have tried using 1mM DDM in 20mM Tris, 5mM
We are trying to purify a membrane protein using different detergents (DDM,
OG etc.). We have tried using 1mM DDM in 20mM Tris, 5mM NaCl and 5%
glycerol buffer to purify the protein. however, we are facing problems in
running the buffer in 16/60 Superdex 200 pg gel filytration coloumn using
AKTA Ex
