Yes, slight overkill.
But I would be concerned about vibrations. Also, would use cabinets in a
coldroom to shield the trays/drops from turbulence created by fans. A
colleague recently told me about excessive nucleation observed when a
plate was left in the coldroom on a bench as opposed to being s
Very nice! You did much leg work...Thanks.
> Folks:
>
> While attending the ICSG meeting in Oxford last month it struck me that
> there might be a need for a general protein crystallography portal
> containing information about software, websites, jobs, journals,
> organizations, education, etc.
Hi Bill,
I would recommend:
Jencks. Catalysis in Chemistry and Enzymology.
Good luck. Maybe in the future I can borrow lecture notes?
Daniel
> Hi Folks:
>
> I just found out that in a couple of weeks I am going to be teaching a
> graduate student-level enzymology course.
>
> Can anyone recom
Hi Alex,
Such gels were used to separate in the past 70S ribosome, plus/minus one
protein - so separation is excellent. Cannot recall the references but
papers published in the 70s' I believe.
The agaose is used to provide "strength" to low percentage acrylamide, say
3% acrylamide/0.5% agarose. J
Dear Isabel,
It worked for a complex that I tested, but crystals were grown in similar
conditions by vapour diffusion using Limbro plates. Advantage was that
small amount of sample was needed. However, one can screen 96 conditions
with as little as ~10 ul of sample using a drop dispensing robot.
Shivesh
If you have not already, maybe try the following:
1. Adding other alcohols as an additive. See work by Keichi Namba in
improving his flagella crystals. The idea is to poison growth in one
direction. I have also had success in "thickening" crystals grown in MPD,
by addition of 1-3% isopro
Best of all, run it on a gel.
> I am pretty sure it is just methylene blue.
>
> You can also see what kind of diffraction pattern the crystal has, or
> just squish it and see if it crumbles.
>
>
> On Jun 14, 2008, at 1:51 PM, Mark Del Campo wrote:
>
>> Before I place an order for some Izit, are th
I would try the following:
1. Remove the His tag.
2. If you need to get rid of the imidazole I would try adding ammonium
sulfate.
3. Is your protein proline- and/or rich in aromatic residues? This may
explain your need for imidazole and solubility.
> Hi there,
>
> Sorry for the off topic questi
Dear Sajid,
I have observed a consistent reduction in mosaicity (from approx. 0.95 to
0.45) for one case by freezing the crystals in a cryostream (N2 vapour) as
opposed to submerging in liquid nitrogen.
Have others observed this?
Daniel
>
> Dear All
>
> My protein size is ~30kD and crystallizes
