Dear all,
I need suggestions regarding phase separation. Upon adding NaCl (100mM) in
the crystallization mixture, the full-fledged oiling is observed after 2
days at 4 degree with long needles (pH 3.6). The Protein contains 50mM of
NaCl. What are the general procedures to get the crystals from oil
Hi James,
I would first ask you what is your crystallization condition?
You mentioned MPD at 70%. Very high concentration (and this is very high) of
organic non volatile alcohols like MPD or polymers like PEGs will promote
phase separation if there is some salt around. The higher the polymer
concen
Dear all,
using ccp4 6.2.1 I have difficulties with the stereochemistry of
oligosaccharide ligands as years ago(My Mail from 2005: stereoisomer as
alternative conformations). REFMAC uses NAG-B-D but we have also NAG-A-D with
an occupancy 0.70:0.30 bound in the same place. REFMAC has problems wi
Dear CCP4bbers,
Surprisingly, only At pH 3.6 the needles are appearing after 3 days but
they are very thin and long rods. I have checked it running SDS-PAGE as well
as using IZIT dye . Since the proteins has already 50mM of NaCl so I added
100mM of NaCl in the crystallization mix. The crystal app
Going from 4 to 25 degrees should let you have your crystals faster (if the
protein stands that temperature, you should have checked it already). As to
improving the crystals, why don't you try the additive kits (several vendors
offer these, including some in a form suited for screening by robot
What do you define as thin and long rods, for some of us this could be
huge crystals. What's your best guestimate regarding the size ? One
way of approximating this is taking a loop of know size and focus on
it, then focus on the crystal and guess.
Small crystals does not necessarily mean the
Hi James,
just out of curiosity. Did IZIT convincingly color the crystals? We have
recently seen that crystals grown in high concentrations of MPD do not color
with IZIT. This phenomenon was documented in Eckert et al. Acta Cryst.
(2003). D59, 139-141.
Best regards
Savvas
Savvas
Hi there,
Would there be a kind soul that could pass this message on to cnsbb? I
am registered with e-mail address velli...@ibs.fr but the e-mail address
has changed to frederic.velli...@ibs.fr 12 months ago perhaps. I am not
very good at all these e-mail diffusion lists and cannot see how to
