[ccp4bb] How to remove nucleic acid contamination for crystallizing zinc finger protein

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From: Tim Fenn fenn {- at -} STANFORD {- dot -} EDU
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Subject: How to remove nucleic acid contamination for crystallizing zinc finger protein
From: tony_htc {- at -} 126 {- dot -} COM tony_htc {- at -} 126 {- dot -} COM
Date: 2007-07-05

Dear all,

Sorry for the off-topic question.

I am purifying a zinc finger transcription factor for crystallization. The protein appeared as a single band on SDS-PAGE (MW 44KD) after NTA chelating column, but its OD280/OD260 ratio is as high as 1.0. So I doubt the protein is nucleic acid contaminated, probably because of the zinc finger. I tried to remove the nucleic acid by Mono Q and Superdex 75 pg, but failed. So could any one recommend some method to remove the nucleic acid during protein crystallization, esp. zinc finger protein? Any experience or references will be appreciated.

Thanks a lot!

Tiancen Hu
Shanghai Institute of Materia Medica

CCP4bb navigation

CCP4bb <-- 2007 <-- July 2007 <-- 05 July 2007

Previous message:
Subject: Re: The CCP4 license is ambiguous
From: Tim Fenn fenn {- at -} STANFORD {- dot -} EDU
Date: 2007-07-05

Next message:
Subject: Re: Off topic: where to order just miniprep columns?
From: Cynthia Kinsland clk10 {- at -} CORNELL {- dot -} EDU
Date: 2007-07-05