Cooling under high pressure reduces the need for cryoprotectant.
http://www.macchess.cornell.edu/MacCHESS/hpcooling.html
On 08/22/16 09:47, Mohammad Saif wrote:
> Dear All,
>
> Would appreciate your suggestions on this. We have solved the
> structure of the enzyme. Now i am trying to crystallize this protein
> with different ligands to study its binding property. First
> established its binding affinity (Kd uM to nM for different ligands).
>
> Co-crystallization didnt work so I tried to seed and soak my enzyme
> crystals into the ligands.
> Soaking I did for 5 mins maximum, as I could see cracks developing in
> the crystal. This was for nM affinity ligand, its stock was made in DMSO.
>
> After processing the data both from seeding and soaking (resolution
> 1.75A to 2.5A), I cannot see the ligand density.
> Any suggestions on what else can be done to get the enzyme- ligand
> crystals.
>
> I see ice-rings in my diffraction images. For cryo-protectant, I use
> mother liquor with 30% glycerol. Shall I increase to say 40% or any
> else salt you would suggest.
>
> Thanks and regards,
> Saif
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All Things Serve the Beam
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David J. Schuller
modern man in a post-modern world
MacCHESS, Cornell University
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