Urea can modify your protein. Maybe you should also try guanidine HCl. Also,
did you try any detergents? Good luck! Jan
--- Sanjiv Kumar <sanjivi...@gmail.com> schrieb am Di, 5.5.2009:
Von: Sanjiv Kumar <sanjivi...@gmail.com>
Betreff: [ccp4bb] Refolding of Denatured Protein
An: CCP4BB@JISCMAIL.AC.UK
Datum: Dienstag, 5. Mai 2009, 13:27
I am working on a membrane protein. There was problem with the normal
purification of the protein so I have tired to purify it under
denaturing conditions (Using 8M Urea). Luckily I could purify the
protein in large quantity. But now the problem is that I am not able to
refold the protein by step wise removing 8M Urea by dialysis (8M, 6M,
4M, 2M, 1M, 0.5M and 0M urea). It is showing aggregation at very high
urea concentration say 2M urea. Kindly suggest any alternate method that I can
try for
refolding of this protein. It is a prokaryotic protein, cloned in
pET28a and expressed in BL21 DE3. Please Help.
Sanjiv Kumar
