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.
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> 1. Re: QUERY (Dr Engelbert Buxbaum)
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> Message: 1
> Date: Tue, 08 Sep 2009 09:58:37 -0400
> From: "Dr Engelbert Buxbaum" <
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> Subject: [Protein-analysis] Re: QUERY
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> Am 04.09.2009, 10:59 Uhr, schrieb Monica Mittal <
monicamittal41@...>:
>
> > 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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