its not going into pellet due to hydrophobicity. It is forming inclusion bodies during overexpression.
You are required to dissolve the protein and refold it.
Please fiollow the procedure "how to refold protein from inclusion bodies"
Good luck with your work
-- Original Message --
From: Monica Mittal <monicamittal41 from yahoo.com>
To: proteins from magpie.bio.indiana.edu
Date: Fri, 4 Sep 2009 20:29:26 +0530 (IST)
Subject: [Protein-analysis] QUERY
I'M FACING PROBLEM IN PURIFYING A protein which is a dna binding protein but its showing a kind of hydrophobic behaviour as its max part is going into pellet after lysis.
moreover its not responding to ni2+ nta column also and the lysis sol. is not so clear that i go for gel filteration as it may probably clog it
so can u suggest any solution?
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