[3dem] protein complex crosslinking for EM

Florian Hauer fhauer at gwdg.de
Tue Jul 1 02:39:14 PDT 2008


Dear Alexandra,

if you want to stabilize your complex by crosslinking, we recommend the  
GraFix method: GraFix: sample preparation for single-particle electron  
cryomicroscopy. Kastner B. et al, Nat Methods. 2008 Jan;5(1):53-5. Epub  
2007 Dec 23.

Simply crosslinking your protein in 'batch modus' will lead to aggregate  
formation. If you have any questions concerning the GraFix protocol, do  
not hesitate to contact me personally.

All the best with your experiments,

Florian.


On Tue, 01 Jul 2008 07:49:33 +0200, Alexandra Deaconescu  
<deacona at brandeis.edu> wrote:

> Dear EM enthusiasts:
>
> I am working on a protein complex that appears to dissociate upon
> dilution for EM grid preparation. I was therefore thinking of using
> some sort of heterobifunctional crosslinker (something other than
> glutaraldehyde that would probably generate a big mess) to stabilize
> the complex and then purify it by gel filtration. I was wondering if
> anyone has and would like to share his/her experiences, or point me
> to some good papers where this has been done before...
>
> I would greatly appreciate your help.
>
> Best,
> Alexandra



-- 
Florian Hauer

3D Cryo Electron Microscopy Group
Max-Planck Institute for biophysical Chemistry
Am Faßberg 11
37077 Göttingen
Germany

Phone: +49 551 201 1302
Fax:   +49 551 201 1197


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