Thanks for all the replies.
I will try a couple of different plates/set-ups. My favorite will be the one
that gives me a crystal ;)
Ø in 2004. Of the 1000 entries that listed [protein], 46 proteins were
crystallized below 3.1 mg/ml.
That is not necessarily the success rate for low concentrations, which we
actually would like to have. We would need negatives for < 3 for to give a
correct answer. I guess even occurrence
Hi,
We have a case where a 260 KDa protein crystallized at 3 mg/ml, that was the
highest concentration we could achieve, without playing with the protein buffer.
Protein-to-well-solution ratio was 3:1.
When screening, we tested all ratios between 4:1 and 1:1. Crystals only
appeared at 3:1 ratio.
We did some data mining from remark 280 of the PDB in 2004. Of the 1000
entries that listed [protein], 46 proteins were crystallized below 3.1
mg/ml. See table 3 at http://www.douglas.co.uk/PDB_data.htm .
Patrick
On 27 February 2012 16:25, Bernhard Rupp (Hofkristallrat a.D.) <
hofkristall...
Hi Sangeetha:
If you just want to check which buffer is good for your protein, maybe you can
try to set up a crystallization screen, keeping your protein concentration just
3 mg/ml. You can observe (after several days) which conditions give you a clear
drop, and maybe you can find a clue which
Quote"Particulalrly the monoolein and monoolein/cholesterol coated plates
( I am not sure I can mention the vendor here but it "should" not matter)"
Since the person who asked this question here
forget about it alltogether to write something back
here is what he was asking about (i think)
Anybody
Yuri,
Did you mean plates for setting up Lipidic Mesophases? If so, here is a
listing of products I have used in the past. I highly endorse the plates
from Molecular dimensions, particularly the plastic laminex plates
(MD11-51-100 + MD11-54). They do have the dis-advantage of drying out
after a
Hi all,
I'm building a ~1.9Å structure that has a few Mg++ ions bound. I thought that
the expected distance for Mg-O was 2.1Å, but in refmac the default to Asp/Glu
oxygens appears to be 1.91 Å. Strangely, for a Mg-bound pyruvate ligand, the
default distance to one oxygen is 2.18 Å, but 1.91 Å f
Why, in the first place, do you feel an urge to concentrate your protein
above 3 mg/ml ?
For crystallization, the concentration needs to be
a) high enough to achieve supersaturation, meaning close enough to the
maximum solubility in a given buffer so that the precipitant can drive the
s
Dear All:
I am having trouble with Coot.
The program keeps crashing when I click on "rotamer analysis". Other
functions, sych as "geometry analysis" all worked fine.
It runs normal before, and only happened when I added the ligands into the
model.
I am using WinCoot_0.7_pre-1-revision-3772, and
I am trying to crystallize a ~320 kDa protein that crashes out if
concentrated past about 3 mg/mL.
I would like to try to exchange it into various buffer-salt-additive
combinations to see which buffer works. For a starting point, I'd like to
use desalting colums.
Does anyone have suggestions
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Dear Sangeetha,
provided you express the protein in E. coli, you could also sonicate the
cell debris in various buffers and compare supernatant/pellet on
SDS-gel. It might already give you a first clue and is faster, cheaper
and you don't risk to clog
Dear bb users,
I am trying to crystallize a ~320 kDa protein that crashes out if
concentrated past about 3 mg/mL.
I would like to try to exchange it into various buffer-salt-additive
combinations to see which buffer works. For a starting point, I'd like to
use desalting colums.
Does anyone have
Dear B,Yes its working. Thanks for your help.
ThanksYogi
> Date: Sun, 26 Feb 2012 22:49:08 +0100
> From: bernh...@chem.gla.ac.uk
> Subject: Re: [ccp4bb] coot with probe and reduce
> To: CCP4BB@JISCMAIL.AC.UK
>
> Dear Raj,
>
> please follow exactly what is written in the FAQ
> http://www.ysbl.
Hi
i do not want to get into trouble by going against any products
The plates what you are talking about as soon it came out we tested.
I did not look back to see if there was a monoolein:cholestrol coated plate.
The ones i used were for sure monoolein coated ones.
That tells it all.The rest of t
Apologies - the limiting date for PhD title is different, as stated here:
The applicant has received PhD degree after March 28, 2008.
Jan Dohnalek
IMC Prague
On Sun, Feb 26, 2012 at 9:52 PM, Jan Dohnalek wrote:
> DEADLINE APPROACHING!
> Our group has been involved in structural studies of sev
We have been doing a bit of this using a simple program under Linux -
pdbskim.
A linux binary available on request. Very simple output though, focused on
infromation on occ, Bs alternatives, etc to help make decisions during
structure refinement.
Jan
On Sat, Feb 25, 2012 at 11:59 PM, WENHE ZHONG
Yuri
I know this isn't quite what you're asking, but it's helpful to use the COC
("UV permeable") version of plates if you're crystallizing samples that
contain detergent. It's just that the drops tend to spread very thinly if
you use the normal polystyrene PS plates. The COC is more hydrophobic
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