Something else you should always try is room-temperature diffraction.
Preferably in-situ diffraction if you can swing it. This is because you
never know if your crystal was "born ugly", or was once beautiful and
peacefully well-ordered before you began batting it around with a nylon
loop lik
Wow! Unexpected advices! Thank you very much, for addon and Fab!
Ed, you are right in both cases: about my name and protease. I think it is
good idea to try to screen for crystallization conditions with bME.
Mahesh, I know nothing about your object, as well as buffer conditions. May
be you would t
Hi Evgeny
I do have a few free cysteine residues but i am not sure if this is the
problem in my case, I say that because i have crystallized this protein
with another ligand without any issues and if oxidation of cysteine
residues is a problem, i would imagine that it would have happened in the
ca
It is very similar with my situation: we are trying to crystallize Fab
fragments. All we got is spherulites. After rMMS-seeding we got better
shaped crystals with only 5 A resolution. So now I think that this
caused by free cysteine near Fab-Fc knee. Oxidation of them leads to
uncontrolled agg
Hi Jürgen
you are right, I did not try any major optimization yet. I only tried to
vary PEG and protein concentration. That did not really improve things too
much. The protein mostly forms spherulites beyond 25% PEG. I am also
thinking that these crystals are poorly diffracting/not diffracting as
Well said Petri,
also how much PEG3350 do you have in your conditions ? More than 25% ? I'm
going after cryo-conditions at this point, you might want to replace your
PEG3350 with smaller PEGs or a mixture of PEG400 and PEG3350.
Almost sounds as if no optimization of the original conditions was p
Hi Petri
They are non-diffracting at the home source and they are cryo cooled. Like
david suggested I guess ill try introducing a buffer as my condition does
not have a buffer. it is ammonium acetate and PEG 3350.
Thanks for the encouragement !
Mahesh
On Mon, Aug 19, 2013 at 5:37 PM, Petri Kur
On 08/19/13 16:49, Mahesh Lingaraju wrote:
Hello people
I recently obtained hexagonal rod like crystals (150x50x20 um) which
turned out to be non diffracting. What is the usual convention for
cases like this ? do people usually give up on the condition or still
try to optimize it ?
It depen
Hello people
I recently obtained hexagonal rod like crystals (150x50x20 um) which turned
out to be non diffracting. What is the usual convention for cases like this
? do people usually give up on the condition or still try to optimize it ?
The crystals are also not very reproducible. I believe it
