grEST purification

Dr. Duncan Clark Duncan at
Fri Mar 3 05:17:57 EST 2000

In article <38C049FB.3207 at>, chenggy
<chenggy at> writes
>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?

Still stuck on the column? Try adding in 20% ethylene glycol to your
buffers or drop down to zero salt and then elute with an increasing urea
gradient i.e. 0 - 4M in Tris buffer plus 20% ethylene glycol.

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382

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