Hello,
While concentrating my GST-tagged protein in a Merck 10kDa cutoff Centricon , the protein appears to be aggregating/precipitating. As is understandable when there is turbidity/whitish appearance in the column . Just simple mixing is just not helping.There is huge protein loss. The buffer I am using for buffer exchange has 50mM Tris-Cl (ph8) and 50mM NaCl. And I have kept the NaCl conc in lysis and washing buffer(for GST purification) at 200mM.
Am I doing the process wrong?
Any suggestions would be appreciated!