K] On Behalf Of Eva
Bligt-Lindén
Sent: 07 November 2012 12:07
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Problems in crystallization
Dear ccp4 users,
I have a problem in the crystallization of my target protein. Whenever
I set up a vapour diffusion experiment, either hanging or sitting
drops, the dr
On Wed, 2012-11-07 at 16:04 +0200, Eva Bligt-Lindén wrote:
> To my knowledge the protein is not a
> surface-tension-reducing-protein and there is no detergent in the
> sample.
However your observations indicate that your sample has reduced surface
tension. When you say that "to your knowledg
Eva
Yes batch under oil will solve the problem. We regularly used to
crystallize conditions with high levels of MPD under oil without difficulty.
I agree that the protein might be reducing the surface tension, but I've
never heard of such a strong effect. Maybe the protein is unstable and
"want
Dear Eva,
are you using siliconized cover slips or normal ones ? Protein drops on not
siliconized glass tend to spread especially if they contain alcohol or
detergent. But this shouldn't be a problem with sitting drops usually.
Good luck !
Ulrike
these xtal tools.
Good luck!
Bryan
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Rob
Gillespie
Sent: Wednesday, November 07, 2012 9:19 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Problems in crystallization
Qiagen used to sell hanging-drop trays with screw
Qiagen used to sell hanging-drop trays with screw-cap tops, and the tops
had six rings on them (3 large, 3 small). The rings had a lip on them that
kept the drop from spreading beyond the edge. I found these quite useful
when setting up drops that had detergent in them.
I unfortunately don't kno
Thank you for your replies. To my knowledge the protein is not a
surface-tension-reducing-protein and there is no detergent in the
sample. I have tried different plates and still the same result. I
will try a different crystallization technique such as batch under oil
or counter-diffusion i
Why don't you try batch under oil?
2012/11/7 Eva Bligt-Lindén
> Dear ccp4 users,
>
> I have a problem in the crystallization of my target protein. Whenever I
> set up a vapour diffusion experiment, either hanging or sitting drops, the
> drops spread out. The surface tension is completely lost in
Hi there,
You could try to go for liquid-liquid interface crystallisation (in
capillaries). The minimum volume would be no less than 1.5 microl
protein + 1.5 microl precipitant, i.e. not the volumes used by nanodrop
robots. Crystallisation under oil (or similar liquids) should also have
the
is your protein a STRP (surface-tension-reducing protein)?
Seriously, did you try different plates? Old-fashioned siliconised cover slips
on Linbro plates?
Does your sample contain detergent?
Mark J van Raaij
Laboratorio M-4
Dpto de Estructura de Macromoleculas
Centro Nacional de Biotecnologia
Dear ccp4 users,
I have a problem in the crystallization of my target protein. Whenever
I set up a vapour diffusion experiment, either hanging or sitting
drops, the drops spread out. The surface tension is completely lost in
80-90% of the droplets. Have any one experienced something similar
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