[ccp4bb] Heavy atom soaking

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CCP4bb <-- 2008 <-- February 2008 <-- 15 February 2008

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Subject: Re: crashing-out protein eluted from Nickel column
From: Antony Oliver Antony {- dot -} Oliver {- at -} ICR {- dot -} AC {- dot -} UK
Date: 2008-02-15

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Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-02-15

Subject: Heavy atom soaking
From: john peter jpeter1000 {- at -} GMAIL {- dot -} COM
Date: 2008-02-15

Dear Folks,
I recently grew crystals in the presence of 100 mM Imidazole pH 8.0 and 1
M Ammonium Phosphate. The protein was in 25 mM Hepes pH 7.4, 10 mM EDTA, 10
mM EGTA and 100 mM NaCl. I like to make heavy atom derivatives with Hg, Pt,
Sm, Au and Pb compounds. I think the imidazole will interfere with Pt.
EDTA and EGTA may also interfere with cations. However I'm not sure about
the effect of phosphate. I tried to transfer the crystal to PEGs by
gradually decreasing the AmPO4 concentration and increasing the PEG
concentration, but the crystals deteriorate. I'm wondering what successful
strategy people have employed to make good heavy atom derivatives when the
mother liquor has salt as the major precipitant. All suggestions, tips are
greatly appreciated. Thanks a lot.

John

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 15 February 2008

Previous message:
Subject: Re: crashing-out protein eluted from Nickel column
From: Antony Oliver Antony {- dot -} Oliver {- at -} ICR {- dot -} AC {- dot -} UK
Date: 2008-02-15

Next message:
Subject: Re: rescuing crashing-out protein eluted from Nickel column
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-02-15