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Re: [ccp4bb] Crosslinking reagents
 

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CCP4bb <-- 2008 <-- March 2008 <-- 04 March 2008
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From: yang li robertcatrukie {- at -} GMAIL {- dot -} COM
Date: 2008-03-04 		Next message:
Subject: Re: phenix.refine and refmac
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Date: 2008-03-04


Subject: Re: Crosslinking reagents
From: David M Shechner shechner {- at -} MIT {- dot -} EDU
Date: 2008-03-04
Quoting Nian Huang :

> I will try that. But I don't think people can tell the final
> concentration of glutaraldehyde in the drop by using its vapor. Maybe
> using direct soaking is what I should do. By the way, should I quench
> it by ammonium or not?
>
> Nian Huang
> Dept of Biochemistry
> UT Southwestern Medical Center
> Dallas, TX 75390

Often it's the case that explicit knowledge of the gluteraldehyde
concentration
in the vapour isn't as important as empirical determination of the exposure
time you subject the drop to, prior to moving it to a new well of
non-crosslinking mother liquor. Here's the first (to my knowledge) reference
that uses this technique; they mention using a time-course to determine
when to
quench:

Lusty, CJ (1999) A gentle vapor-diffusion technique for cross-linking
of protein
crystals for cryocrystallography. J. Appl. Cryst. 32. 106-112.

Cheers,
d.s.

===========================================
David M. Shechner, Graduate Student
Bartel Laboratory
Department of Biology
Whitehead Institute for Biomedical Research
Massachusetts Institute of Technology
shechner@mit.edu
===========================================
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