his purification

UTHANDI,SIVAKUMAR via methods%40net.bio.net (by usiva from ufl.edu)
Tue Jun 8 14:12:25 EST 2010

Hi All
I am trying to purify His-tagged protein from a halophilic (2M 
NaCl)archaea which is around 30kDa. I was succesful in purifying 
thro nickel column. When I run the His purified protein sample 
(after dialysis in Tris buffer containing 2M NaCl, dilaysis also 
helps to remove excess immidazol from the protein) on to the 
gelfiltration coulumn(superdex 200), it was eluting at a molecular 
weight of ~460kDa. I dont know whether the protein is getting 
complexed or aggregated? But shows single band on SDS Page.
Can any one has the similar problem? Any help please?
Thanks, Siva

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