Re: [ccp4bb] question about the zinc binding protein
Hi, I'm working with a possible zinc binding protein. Saw your post and was wondering, what is the proper pH range for zinc binding? Thanks, Peter
Re: [ccp4bb] question about the zinc binding protein
I have no idea, it is not my buffer, it is dengzq1987's! 'BTW' means 'by the way', BTW. cheers, charlie >>> Enrico Stura 4/1/2010 11:01 am >>> What is 1mM BTW? I am not familiar with this abbreviation. PBS phoshate buffered saline (Phosphate + NaCl) not suitable for zinc binding proteins BBS borate buffered saline (Borate + NaCl) wrong pH for zinc binding. BTW ? (? ? ?) No idea what this is. Charles, do you know if it includes HEPES or Phosphate? Enrico. On Thu, 01 Apr 2010 11:06:10 +0200, Charles Allerston wrote: > Hi, > > are you sure it is your protein precipitating? You will get a cloudy > precipitate appearing in HEPES and Phosphate buffers on addition of > ZnCl, without protein. > > > cheers > > charlie > dengzq1987 3/31/2010 5:08 pm >>> > hello everyone, recently i purify a protein conteining zinc binding > domain,and i want to determine its structure.i get the crystal,but poor > diffraction.so i try to adding zinc into the protein to optimize the > crystal,but the protein precipitate immidiately even the znic is 1 > mM.BTW,we use the protein to do zinc scan,we don't find the zinc. does > anyone have some advice? > > 2010-03-31 > > > > dengzq1987 -- Enrico A. Stura D.Phil. (Oxon) ,Tel: 33 (0)1 69 08 4302 Office Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab LTMB, SIMOPRO, IBiTec-S, CEA Saclay, 91191 Gif-sur-Yvette Cedex FRANCE http://www-dsv.cea.fr/en/ibitecs/82 http://www.chem.gla.ac.uk/protein/mirror/stura/index2.html e-mail: est...@cea.fr Fax: 33 (0)1 69 08 90 71
Re: [ccp4bb] question about the zinc binding protein
BTW"By the way" I think, not a buffer abbreviation -- AstraZeneca UK Limited is a company incorporated in England and Wales with registered number: 03674842 and a registered office at 15 Stanhope Gate, London W1K 1LN. Confidentiality Notice: This message is private and may contain confidential, proprietary and legally privileged information. If you have received this message in error, please notify us and remove it from your system and note that you must not copy, distribute or take any action in reliance on it. Any unauthorised use or disclosure of the contents of this message is not permitted and may be unlawful. Disclaimer: Email messages may be subject to delays, interception, non-delivery and unauthorised alterations. Therefore, information expressed in this message is not given or endorsed by AstraZeneca UK Limited unless otherwise notified by an authorised representative independent of this message. No contractual relationship is created by this message by any person unless specifically indicated by agreement in writing other than email. Monitoring: AstraZeneca UK Limited may monitor email traffic data and content for the purposes of the prevention and detection of crime, ensuring the security of our computer systems and checking Compliance with our Code of Conduct and Policies. -Original Message- From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Enrico Stura Sent: 01 April 2010 11:02 To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] question about the zinc binding protein What is 1mM BTW? I am not familiar with this abbreviation. PBS phoshate buffered saline (Phosphate + NaCl) not suitable for zinc binding proteins BBS borate buffered saline (Borate + NaCl) wrong pH for zinc binding. BTW ? (? ? ?) No idea what this is. Charles, do you know if it includes HEPES or Phosphate? Enrico. On Thu, 01 Apr 2010 11:06:10 +0200, Charles Allerston wrote: > Hi, > > are you sure it is your protein precipitating? You will get a cloudy > precipitate appearing in HEPES and Phosphate buffers on addition of > ZnCl, without protein. > > > cheers > > charlie > >>>> dengzq1987 3/31/2010 5:08 pm >>> > hello everyone, recently i purify a protein conteining zinc binding > domain,and i want to determine its structure.i get the crystal,but poor > diffraction.so i try to adding zinc into the protein to optimize the > crystal,but the protein precipitate immidiately even the znic is 1 > mM.BTW,we use the protein to do zinc scan,we don't find the zinc. does > anyone have some advice? > > 2010-03-31 > > > > dengzq1987 -- Enrico A. Stura D.Phil. (Oxon) ,Tel: 33 (0)1 69 08 4302 Office Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab LTMB, SIMOPRO, IBiTec-S, CEA Saclay, 91191 Gif-sur-Yvette Cedex FRANCE http://www-dsv.cea.fr/en/ibitecs/82 http://www.chem.gla.ac.uk/protein/mirror/stura/index2.html e-mail: est...@cea.fr Fax: 33 (0)1 69 08 90 71
Re: [ccp4bb] question about the zinc binding protein
What is 1mM BTW? I am not familiar with this abbreviation. PBS phoshate buffered saline (Phosphate + NaCl) not suitable for zinc binding proteins BBS borate buffered saline (Borate + NaCl) wrong pH for zinc binding. BTW ? (? ? ?) No idea what this is. Charles, do you know if it includes HEPES or Phosphate? Enrico. On Thu, 01 Apr 2010 11:06:10 +0200, Charles Allerston wrote: Hi, are you sure it is your protein precipitating? You will get a cloudy precipitate appearing in HEPES and Phosphate buffers on addition of ZnCl, without protein. cheers charlie dengzq1987 3/31/2010 5:08 pm >>> hello everyone, recently i purify a protein conteining zinc binding domain,and i want to determine its structure.i get the crystal,but poor diffraction.so i try to adding zinc into the protein to optimize the crystal,but the protein precipitate immidiately even the znic is 1 mM.BTW,we use the protein to do zinc scan,we don't find the zinc. does anyone have some advice? 2010-03-31 dengzq1987 -- Enrico A. Stura D.Phil. (Oxon) ,Tel: 33 (0)1 69 08 4302 Office Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab LTMB, SIMOPRO, IBiTec-S, CEA Saclay, 91191 Gif-sur-Yvette Cedex FRANCE http://www-dsv.cea.fr/en/ibitecs/82 http://www.chem.gla.ac.uk/protein/mirror/stura/index2.html e-mail: est...@cea.fr Fax: 33 (0)1 69 08 90 71
Re: [ccp4bb] question about the zinc binding protein
Hi, are you sure it is your protein precipitating? You will get a cloudy precipitate appearing in HEPES and Phosphate buffers on addition of ZnCl, without protein. cheers charlie >>> dengzq1987 3/31/2010 5:08 pm >>> hello everyone, recently i purify a protein conteining zinc binding domain,and i want to determine its structure.i get the crystal,but poor diffraction.so i try to adding zinc into the protein to optimize the crystal,but the protein precipitate immidiately even the znic is 1 mM.BTW,we use the protein to do zinc scan,we don't find the zinc. does anyone have some advice? 2010-03-31 dengzq1987
