leela shetty wants you to join Yaari!
Is leela your friend?
Please respond or leela may think you said no :(
This being Valentine's Day and all, am I the only one who is wondering if
Leela and CCP4BB hit it off?
--dvd
**
Hi all,
I have protein crystals in complex with substrate grown in 20-30% (w/
v) PEG 3350, 4% (w/v) ammonium sulfate, and 0.1M sodium MES buffer at
pH 6.5. I purify and concentrate my protein in a high salt buffer
(0.5M NaCl, 0.1M sodium MES at pH 6.5, 10% glycerol, 2mM DTT).
I grow my
Keith,
There could be as many number of stories of cryoprotection to as many number of
structures deposited in the pdb.
PEG3350 seems to be little too large for freezing, particularly in presence of
so much
of salt. I have collected data up to 1.0 Ang with PEG2000 freezing.
If you have not
Hi Keith,
I think there is a lot of advice out there on rational design of the
cryoprotectant. One of the most useful in your case is one that I think
originates with Elspeth Garman (but she will correct me if I'm wrong).
This is osmolarity matching. I don't have a reference to hand but you can
Hi Keith,
PEG3350 freezes ~30% in combination with 10% glycerol, since you
already have some glycerol from your protein I would increase both PEG
and glycerol say 30% PEG3350 and 20% glycerol - but you tried this
already.
Your real problem is the big crystal, you might want to consider
