Are you sure the aggregation is not caused by dna interweaving between protein molecules? If that's the cause, Benzonase will be a good choice for DNA clearance.
> -----Original E-mail-----
> From: proteins-request from oat.bio.indiana.edu> Sent Time: 2009-9-10 1:04:09
> To: proteins from magpie.bio.indiana.edu> Cc:
> Subject: Proteins Digest, Vol 52, Issue 3
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>>> Today's Topics:
>> 1. Re: QUERY (Dr Engelbert Buxbaum)
>>> ----------------------------------------------------------------------
>> Message: 1
> Date: Tue, 08 Sep 2009 09:58:37 -0400
> From: "Dr Engelbert Buxbaum" <engelbert_buxbaum from hotmail.com>
> Subject: [Protein-analysis] Re: QUERY
> To: proteins from net.bio.net> Message-ID: <op.uzxyfzdu66vu6s from bengelbert-dm.rusm.rossu.loc>
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>> Am 04.09.2009, 10:59 Uhr, schrieb Monica Mittal <monicamittal41 from yahoo.com>:
>> > HI
> > 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?
>> Does that mean you are expressing a poly-His tagged protein in bacteria?
> If so, have you checked that your protein is not in inclusion bodies? How
> did you lyse? How long and at what g did you try to clarify your lysate (1
> h at 100000 g should do the trick)? Have you fragmented the bacterial DNA,
> and are you sure that your protein is not binding to the fragments? Are
> you protecting against oxidation and proteolysis?
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