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
