[Protein-analysis] Re: Proteins Digest, Vol 52, Issue 3

winlight from mail.ustc.edu.cn via proteins%40net.bio.net (by winlight from mail.ustc.edu.cn)
Sun Sep 13 20:22:25 EST 2009


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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> Subject: Proteins Digest, Vol 52, Issue 3
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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" <engelbert_buxbaum from hotmail.com>
> Subject: [Protein-analysis] Re: QUERY
> To: proteins from net.bio.net
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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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