Dear All,
This question sounds simple but I dont know the answer.I was preparing a 24
well crystal screen. When I try to use 10 mM ZnSO4 with HEPES (pH 7.6) buffer
it precipitates. I tried both ZnCl2 and Zn acetate the effect is same. I dont
know why this Zn in not compatible with
Try adding water first, so that you are not mixing concentrated Zn with
concentrated HEPES. Also it depends what else is in your cocktail.
JPK
On Fri, May 11, 2012 at 11:26 AM, Rajesh Kumar ccp4...@hotmail.com wrote:
Dear All,
This question sounds simple but I dont know the answer.
I was
On 05/11/12 12:26, Rajesh Kumar wrote:
Dear All,
This question sounds simple but I dont know the answer.
I was preparing a 24 well crystal screen. When I try to use 10 mM
ZnSO4 with HEPES (pH 7.6) buffer it precipitates. I tried both ZnCl2
and Zn acetate the effect is same.
I dont know why
many might have used
this combination.
ThanksRajesh
Date: Fri, 11 May 2012 11:29:03 -0500
Subject: Re: [ccp4bb] zinc with HEPES
From: j-kell...@fsm.northwestern.edu
To: ccp4...@hotmail.com
CC: CCP4BB@jiscmail.ac.uk
Try adding water first, so that you are not mixing concentrated Zn
If there is no cure , then fine.pH may not be the answer as it doesn't Happen
with TRIS buffer pH 7.6.Thanks to every oneRajesh
Date: Fri, 11 May 2012 12:35:45 -0400
From: dj...@cornell.edu
Subject: Re: [ccp4bb] zinc with HEPES
To: CCP4BB@JISCMAIL.AC.UK
pH is the culprit here
Like some already mentioned
change your pH to 6.4 and use a different buffer like cacodylate
or you can use Zinc acetate in water pH still would be 6.4
from your last mail
why would you add Zn to your initial hit condition
is there any rationale?
Padayatti
On Fri, May 11,
, then fine.
pH may not be the answer as it doesn't Happen with TRIS buffer pH 7.6.
Thanks to every one
Rajesh
--
Date: Fri, 11 May 2012 12:35:45 -0400
From: dj...@cornell.edu
Subject: Re: [ccp4bb] zinc with HEPES
To: CCP4BB@JISCMAIL.AC.UK
There is no cure
increase
thickness or change the shape of very very thin needle crystals.
Yes, I will try very less, 50uM.Thanks for helping me to understand.Rajesh
Date: Fri, 11 May 2012 12:53:53 -0400
Subject: Re: [ccp4bb] zinc with HEPES
From: liehy...@gmail.com
To: ccp4...@hotmail.com
Rajesh,
10mM zinc
buffer pH 7.6.
Thanks to every one
Rajesh
--
Date: Fri, 11 May 2012 12:35:45 -0400
From: dj...@cornell.edu
Subject: Re: [ccp4bb] zinc with HEPES
To: CCP4BB@JISCMAIL.AC.UK
There is no cure for HEPES
Almost assuredly you are getting zinc hydroxide precipitate. Zn(OH)2
will readily precipitate from solutions of alkaline Zn(II). Your problem
is compounded by the fact that HEPES is a non-coordinating buffer, so it
does not help solubilize the zinc ion. You might find a weakly
coordinating
Date: Fri, 11 May 2012 18:09:54 +0100
Subject: Re: [ccp4bb] zinc with HEPES
From: patr...@douglas.co.uk
To: ccp4...@hotmail.com
Rajesh
How did you do the MMS? By hand or with a robot, and what screens did you use?
and why did you change to HEPES out of interest?
Patrick
On 11 May 2012 18:05, Rajesh
several drops to just to fish out
1-2 needles. I am ready to try if there any trick left.
Thanks for lots and lots of help.
Regards,
Rajesh
--
Date: Fri, 11 May 2012 18:09:54 +0100
Subject: Re: [ccp4bb] zinc with HEPES
From: patr...@douglas.co.uk
To: ccp4
to try if there any trick left.
Thanks for lots and lots of help.
Regards,
Rajesh
--
Date: Fri, 11 May 2012 18:09:54 +0100
Subject: Re: [ccp4bb] zinc with HEPES
From: patr...@douglas.co.uk
To: ccp4...@hotmail.com
Rajesh
How did you do the MMS? By hand
.
Thanks to every one
Rajesh
Date: Fri, 11 May 2012 12:35:45 -0400
From: dj...@cornell.edu mailto:dj...@cornell.edu
Subject: Re: [ccp4bb] zinc with HEPES
To: CCP4BB@JISCMAIL.AC.UK
not be the answer as it doesn't Happen with TRIS buffer pH 7.6.
Thanks to every one
Rajesh
--
Date: Fri, 11 May 2012 12:35:45 -0400
From: dj...@cornell.edu
Subject: Re: [ccp4bb] zinc with HEPES
To: CCP4BB@JISCMAIL.AC.UK
There is no cure for HEPES
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