grEST purification

chenggy chenggy at
Fri Mar 3 02:23:16 EST 2000

Hi, Everyone! 

I am purifying two gravity related esterase, enzyme  are stable
at the present of low concentration of SDS and high resolution 
of (NH4)2SO4, but the activity lost at the present of SDS after
the phenol sepharose column.

In column work, 
   starting and washing buffer : 50 mM Tris-Cl buffer containing
                                    1.7 M (NH4)2SO4  
eluted with gradient (NH4)2SO4  from 1.7 M to 0 M. 

Where is the enzyme? How can I found the reson?

Please send me your idea. 
With best regards. 
Thanks a lot  for your reply...... 

chenggy at


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