DLadakis,
The seed stock solution should be your crystallization solution with a
slight (5% maybe) increase in the precipitant. In my experience, I have not
needed additional protein to stabilize the nano-crystals, but as with all
things, every protein is different.
The idea is the crystals should
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Dear DLadakis,
you are probably thinking of using a robot for seeding. For manual
seeding you would overcome the problem of a stock solution by using a
cat whisker for seeding. Manual seeding also has the advantage of
getting a much better feeling for
Dear all
I would like your advice on preparing a seed stock for microseeding.
Should the seed stabilizing stock soltion include soluble protein as well. If
it doesn't will the seeds eventually disolve leading to an unsuccesfull screen?
Also how is the seedstock supposed to look under the microscop
Patrick,
This thing all things devours:
Birds, beasts, trees, flowers;
Gnaws iron, bites steel;
Grinds hard stones to meal;
Slays king, ruins town
And beats high mountains down.
Answer: TIME
When scaling up in seeding the most important factor is: "TIME"
While the environment may be the
Sorry, I said that last part wrong. I meant it is sometimes helpful to*increase
* the salt by 50% when scaling up.
On 19 March 2012 22:29, Patrick Shaw Stewart wrote:
>
> Rajesh
>
> If you set up the volumes you suggest you will probably get
> precipitation. This is counterintuitive until you
Rajesh
If you set up the volumes you suggest you will probably get precipitation.
This is counterintuitive until you realize that (as Ed says) you will be
losing a lot of protein with those small drops. When you scale up the
surface area to volume ratio is lower, so a smaller proportion of the
pr
Scaling up 100nl drops is problematic. What I understand is that it is
not only the different equilibration conditions, but primarily the
amount of protein that gets absorbed on the surface is relatively higher
for small drops. There were some empirical formula for scaling up (i.e.
how much you n
Dear All,
I have few papers in hand which explain me about microseeding, matrix
microseeding, and cross seeding.I have also read few earlier threads and some
more literature in google.Using Phoenix robot, I did a matrix micro-seeding and
matrix cross seeding. I have few hits with this.In 96 we
See the following publication for an interesting method to resolve
pseudo-merohedral twinning in SeMet crystals:
Cansizoglu, A. E. and Chook, Y. M. (2007) Conformational heterogeneity
of Karyopherinβ2 is segmental. Structure, 15(11):1431-1441.
"In an effort to obtain single selenomethionine
Yes, on both counts. SeMet proteins can be frustratingly* close to
their native forms - with little quirks like twinning or lack of
useful diffraction etc. It's common practice to micro (or macro) seed
with native crystals and it often works quite well. It's fun and
sometimes useful to add a few ul
Dear All,
My SeMet protein crystals have a needle-like morphology, worse than that of
the native xtals. Also, although the cell dimensions of both forms is very
much similar, the SeMet xtals are twinned(as indicated by Ctruncate plots).
I was wondering if such cases were commonly seen, also is it
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