From: K Singh
Date: 29 September 2011 09:54
Dear All,
Apologies for bit off-topic. I am in process of purifying a protein by
ion-exchange (cation)
using 0.5-3M NaCl gradient. And I get a sharp peak which is western
positive. Following elution,
I wash the column >10 column volume with 3M NaCl. and now when I
wash the column
with water, I get a huge peak which is also western positive. Any
inputs or suggestions explaining
this phenomenon?
Kris
----------
From: Roger Rowlett
Date: 29 September 2011 09:54
Dear All,
Apologies for bit off-topic. I am in process of purifying a protein by
ion-exchange (cation)
using 0.5-3M NaCl gradient. And I get a sharp peak which is western
positive. Following elution,
I wash the column >10 column volume with 3M NaCl. and now when I
wash the column
with water, I get a huge peak which is also western positive. Any
inputs or suggestions explaining
this phenomenon?
Kris
----------
From: Roger Rowlett
Since water is unbuffered, you are likely changing the pH of the solvent, desorbing additional protein. If you are working unbuffered, your IEX results will be unpredictable.
Roger Rowlett
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