Dear Sandrine,
On a Gel filtration column you are expected to get the 25 KDa peak
earlier than 15 KDa.
A number of factors affect the resolution, check the length of the
column, amount of load etc. If you can give the details of the run and
column contition it would be easier for someone ( including me, of
course) to answer your query)
Best of Luck
S.Ballal
Indus Biotherapeutics Ltd.
Ahmedabad, India
http://www.indusbio.co.insandrine.vessillier at caramail.com wrote in message news:<20030320153003.9102.qmail at ww02.hostica.com>...
> Hi,
> I actually try to purify 2 proteins by gel filtration, one is 25KDa and the other one 15 KDa. SDS PAGE analysis indicates the first pic to be eluted is the 15KDa protein and I can not see the 25 KDa protein. Do you think it is retained on the column ?(I use 6M guanidine and 0.5M NaCl in tris buffer pH8 for loading and separation). Do you have some suggestions?
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