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Re: [ccp4bb] cryoprotectant for crystals in isopropanol
 

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CCP4bb <-- 2008 <-- March 2008 <-- 19 March 2008
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Subject: Post-Doctoral position at the University of Dundee
From: Paul Fyfe P {- dot -} K {- dot -} Fyfe {- at -} DUNDEE {- dot -} AC {- dot -} UK
Date: 2008-03-19 		Next message:
Subject: PRODRG?
From: "Harry M {- dot -} Greenblatt" Harry {- dot -} Greenblatt {- at -} WEIZMANN {- dot -} AC {- dot -} IL
Date: 2008-03-19


Subject: Re: cryoprotectant for crystals in isopropanol
From: David Roberts droberts {- at -} DEPAUW {- dot -} EDU
Date: 2008-03-19
Not that all proteins are created alike, but we had a protein
crystallize in a similar situation (PEG/20%IPA). I don't remember the
exact details, but we did have good luck using just glycerol as a
cryoprotectant. That is, I made up the well buffer and added to that
well buffer 20-25% glycerol (in addition to IPA/PEG), and moved the
crystal into that. We had no issues with cracking or anything, and data
collection went well. The first thing I tried worked, so I don't have a
lot of trial information for you. You could probably reduce the
glycerol a bit with no worries, but I don't know.

Good luck

Dave

shivesh kumar wrote:
> Dear all,
> The question is regarding the cryoprotectant.We have crystallized a
> protein in 20% PEG 500 with 10% isopropanol.What should be the cryo we
> should try for data collection.Also,what percentage of PEG 500 is
> sufficient enough as a cryo for data collection.
> We are also trying to crystallize it with higher PEG's also like 1K,
> 2K, 4K.Lets C.Since the condition is having isopropanol also and it is
> reported that mounting with isoprop is very difficult and it
> evaporates as soon as we open the slides.
> Thanx in advance...
> Shivesh Kumar

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CCP4bb <-- 2008 <-- March 2008 <-- 19 March 2008
Previous message:
Subject: Post-Doctoral position at the University of Dundee
From: Paul Fyfe P {- dot -} K {- dot -} Fyfe {- at -} DUNDEE {- dot -} AC {- dot -} UK
Date: 2008-03-19 		Next message:
Subject: PRODRG?
From: "Harry M {- dot -} Greenblatt" Harry {- dot -} Greenblatt {- at -} WEIZMANN {- dot -} AC {- dot -} IL
Date: 2008-03-19


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