llization diagram I’d love to hear.
>
>
>
> *Regards,*
>
>
>
> *Vaheh Oganesyan*
>
> *www.medimmune.com <http://www.medimmune.com>*
>
>
>
> *From:* CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *On Behalf Of
> *Philippe
> BENAS
> *Sent:* Thur
Oganesyan, Vaheh
Sent: Thursday, July 13, 2017 9:50 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] crystallization optimization
What I’m about to write should be referred as a question rather than an answer.
However, it might also help to find the answer to crystallization question
discussed
t mailto:patr...@douglas.co.uk>>
À : CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>
Envoyé le : Mercredi 12 juillet 2017 17h28
Objet : Re: [ccp4bb] crystallization optimization
Alun
I agree Frank's point is very interesting - and he intriguingly refers us to
the phase dia
Wales:
Registration number 1794026
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Debanu Das
Sent: 12 July 2017 22:38
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] crystallization optimization
Yes, we have done this (addition of water to dilute screen reagents in the
Yes, we have our screenmaking robot programmed to set 50% diluted screens
and frequently employ this when a large fraction of the undiluted screens
result in precipitation. We have found quite a few productive hits this
way, as some previously grungy wells often present crystals when diluted.
Chee
Yes, we have done this (addition of water to dilute screen reagents in the
well) and also try it now in some cases and in fact, this is also the rationale
behind Hampton's Crystal Screen Lite.
Best,
Debanu
--
Debanu Das
> On Jul 12, 2017, at 9:01 AM, Alun R Coker wrote:
>
> So, if we have a co
t;CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>"
mailto:CCP4BB@JISCMAIL.AC.UK>>
Subject: Re: [ccp4bb] crystallization optimization
Yes, exactly. Thanks for doing the Right Thing and posting the actual diagram.
On 12/07/2017 16:26, Patrick Shaw Stewart wrote:
Alun
I agree
So, if we have a commercial 96 well screen where more than around 60% of
the drops precipitate. It may be worth diluting the whole screen say
(30ul screen and 20ul water in each well) and repeating . rather
than diluting the protein.
Has anyone ever tried this?
All the best,
Alun
On 12
Yes, exactly. Thanks for doing the Right Thing and posting the actual
diagram.
On 12/07/2017 16:26, Patrick Shaw Stewart wrote:
Alun
I agree Frank's point is very interesting - and he intriguingly refers
us to the phase diagram.
Is the point that Line A is longer than Line B ?
Best wish
_
From: CCP4 bulletin board on behalf of Emmanuel
Saridakis
Sent: 12 July 2017 13:55:31
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] crystallization optimization
Dear All,
Fine-tuning protein and precipitant concentration is of course the first line
of approach, followed by both
, 293.
E E . Saridakis and N.E. Chayen. Protein Science (2000) 9 , 755.
Best,
Emmanuel
De: "Alun R Coker"
À: "CCP4BB"
Envoyé: Mercredi 12 Juillet 2017 15:40:10
Objet: Re: [ccp4bb] crystallization optimization
Hi Everyone,
Franks point is really interesting.
Hi Everyone,
Franks point is really interesting. We routinely reduce the protein
concentration when we see too many precipitated wells, but we never
dilute the screen. Has anyone tried this?
All the best,
Alun
On 12/07/17 08:48, Frank von Delft wrote:
The point I was failing to make: re
Dear Patrick,
Of course this is the best-first choice!
I got confused with the names in my previous email. I was reffering to Chen
when I mentioned the screens.
Kindly,
On Wed, Jul 12, 2017 at 12:48 PM, Olga Moroz wrote:
> MMS (microseed matrix screening) into several screens, as Patrick
> sug
MMS (microseed matrix screening) into several screens, as Patrick suggested
earlier, would be the first choice for me.
Works very often.
Good luck!
Olga
> On 12 Jul 2017, at 08:48, Frank von Delft wrote:
>
> The point I was failing to make: reducing either protein or precipitant
> concentr
Vicky, streak seeding is a very good method, but it can be quite a lot of
work. Before he tries that, why wouldn't we suggest to Liuqing that he
should try MMS - that is, adding a liquid seed stock to random screens?
That way he is likely to end up with seeds in wells with similar conditions
that
The point I was failing to make: reducing either protein or precipitant
concentration will indeed reduce nucleation, but often won't get you
bigger or more single crystals: it will just make the appearance of
crystals less reliable.
The way to get big single reliable crystals is to /increase
Dear Frank,
I may see in the attached pic several nucleation points and a considerable
amount of microcrystals. Based to my knowledge decreasing the concentration
of the precipitant and/or the protein concentration would be a reasonable
approach to refine the initial hits.
By checking the diagram
Actually, you should try /increasing/ the protein concentration - a
lot. But be prepared to drop the precipitant concentration to almost
nothing (1 or 2% isn't "low").
To understand why, look at the phase diagram and what we assume about
vapour diffusion. (Which I'm assuming is what you're d
Dear Patrick,
You may reduce the protein concentation, as well.
Another option could be the *streak seeding* by exploiting the drop of your
initial condition.
Good luck,
V.T.
On Mon, Jul 10, 2017 at 7:17 PM, Patrick Shaw Stewart wrote:
>
> Microseed them into two or three random screens.
>
>
Microseed them into two or three random screens.
Search for MMS and rMMS online.
Good luck
Patrick
On 10 July 2017 at 15:47, Liuqing Chen <519198...@163.com> wrote:
> hello everyone!
> I get a condition (10% w/v PEG 6000, 100mm HEPES PH7.0) in which my
> protein grow small needle like crys
hello everyone!
I get a condition (10% w/v PEG 6000, 100mm HEPES PH7.0) in which my protein
grow small needle like crystals, how can i optimize it to get bigger crystals?
the attach is the crystals figure.
thanks in advance
sincerely
Liuqing Chen
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